FR2733995A1 - INHIBITORS OF THE INACTIVATION OF ENDOGENOUS NEUROPEPTIDES, IN PARTICULAR CHOLECYSTOKININ, THEIR METHODS OF PREPARATION THEIR USE AS MEDICINAL PRODUCTS AND METHOD OF SCREENING OF MEDICINES - Google Patents

Abstract

Inhibitors of a membrane tripeptidylpeptidase responsible for the inactivation of endogenous neuropeptides such as cholecystokinin are described.

Description

INHIBITORS OF INACTIVATION OF ENDOGENOUS NEUROPEPTIDES
IN PARTICULAR THE CHOLECYSTOKININE, THEIR METHODS OF PREPARATION,
THEIR USE AS MEDICAMENTS AND METHOD FOR SCREENING
DRUGS
The present invention is in the field of the prevention of inactivation of endogenous neuropeptides such as cholecystokinin (CCK).

 Also, the invention relates more specifically to a process for the pure preparation of enzymes involved in this type of inactivation. It also relates to the nucleotide sequences encoding said enzymes It also relates to the enzymes thus obtained.

 The subject of the invention is also a method for screening medicinal products using said enzymes.

 In addition, the subject of the present invention is chemical compounds of defined formula as well as their processes of preparation and their use as inhibitors of the abovementioned enzymes, involved in the physiological inactivation of endogenous neuropeptides such as, in particular, cholecystokinin (CCK) .

 The subject of the invention is also physiologically acceptable compositions comprising said compounds.

 It also relates to the use of said compounds in the preparation of medicaments for humans or animals.

 The important physiological role of endogenous neuropeptides and in particular that of cholecystokinin (CCK) is well known. The latter is both a digestive hormone and a neurotransmitter of the central and peripheral nervous systems. In the latter case, it is mainly in the form of a sulphated octapeptide (CCK-8-S) corresponding to the formula Asp-Tyr (SO3H) -Met-Gly-Trp-Met-Asp-Phe-NH2.

 CCK has various actions on motility and digestive secretions (contraction of the gallbladder, inhibition of gastric secretion, etc.) as well as on the central nervous system (analgesia, action on mood and cognition) and endocrine (pituitary secretions). In filtering, CCK and certain derivatives possess potent anorexigenic activity by facilitating satiety by stimulating peripheral and possibly central receptors (J.J.

Vanderhaegen and J.M. Crawley, Ann. N.Y. Acad. Sci., 1985, 448: 677).

 Although interesting effects of the pharmacological stimulation of CCK receptors could have been expected, no stimulating agent for these receptors is currently available for therapeutic use mainly for reasons of poor bioavailability of CCK and its derivatives. In addition, it has even been shown that direct stimulation of CCK receptors is likely to cause undesirable anxiogenic effects.

 Other studies have focused on the identification of the enzyme or enzymes responsible for the physiological inactivation of these endogenous neuropeptides and in particular CCK but have not led to a characterization so far. sufficiently precise to allow the development of inhibitors.

Thus it has been suggested the involvement of different enzymes that are supposed to attack certain peptide bonds of the CCK-8 molecule, such as in particular that of an acid aminopeptidase, distinct from aminopeptidase A (Deschodt-Lanckman et al. Peptides, 1983, 4: 71), enkephalinase and aminopeptidase (Matsas et al., FEBS
Lett., 1984, 175: 124 Deschodt-Lanckman et al., Regul.

Peptides, 1981, 2:15), that of thiolpeptidases (Mc Dermott et al., Neurochem Int, 1983, 5: 641 1 Durieux et al.,
Neuropeptides, 1986, 7: 1) as well as that of a metalloendopeptidase (Steardo et al., J. Neurochem., 1985, 45,784).

 More recently, Rose et al. (Proc Natl Acad, Sci USA, 1988, 85: 8326, Neurosci, 1989, 29: 583) have suggested the intervention of a serine peptidase. They further found that general serine group reagents proteins such as diisopropylfluorophosphorate (DFP) or chloromethylketones or boronic acid elastase inhibitors (a family of serine peptidases) were able to prevent the enzymatic degradation of endogenous CCK-8 released by depolarization of brain slices.

 However, these experiments have failed to identify and purify the enzyme responsible and the compounds used in these in vitro experiments can not be considered as drugs, in particular, because of their toxicity, their lack of specificity and / or their poor bioavailability.

 It is therefore found that there remains in the state of the art the need to precisely identify the enzyme or enzymes responsible for the inactivation of endogenous neuropeptides and in particular cholecystokinin (CCK) and the need for non-toxic, bioavailable, specific chemical compounds that can be used as drugs for the prevention of inactivation of these endogenous neuropeptides.

 The invention has now made it possible to achieve these objectives by providing, in particular, a method for screening drugs using the isolated responsible enzyme, thus making it possible, depending on whether said enzyme is inhibited or not, to distinguish molecules that may constitute effective drugs in the treatment of disorders or conditions where endogenous neuropeptides, and in particular cholecystokinin, are involved.

 The present invention also provides chemical compounds of formula defined below, which can be used in the prevention of the inactivation of endogenous neuropeptides and thus satisfies the other essential need remaining in the state of the art.

Thus, the present invention firstly relates to a process for preparing a membrane tripptidylpeptidase in the pure state, characterized in that it comprises the following steps
i) preparation of brain membranes (cerebral cortex), for example rat
ii) purification by high performance liquid chromatography (HPLC)
iii) verification of the product obtained by enzymatic reaction using a CCK substrate, for example the peptides
CCK-8 (non-sulphated) of formula Asp-Tyr-Met-Gly-Trp-Met-Asp-Phe-NH2
or CCK-5 of formula
Gly-Trp-Met-Asp-Phe-NH2
The subject of the invention is also the purified enzyme thus obtained.

 The study of the specificity of this purified enzyme on a series of model substrates has shown that it behaves like an aminotripeptidylpeptidase.

 The inventors have performed sequencing work and have surprisingly found that a fragment of the purified protein has a strong similarity to a protease known as tripeptidyl peptidase II previously isolated from human erythrocytes or rat liver. (Balow RM et al., J. Biol Chem., 1986, 261: 24092417). However, the function of the latter remained totally unknown and its role in the inactivation of neuropeptides, such as CCK, not suspected especially since it was known only as a cytoplasmic enzyme.

Using traditional methods of molecular cloning, the inventors identified two distinct clones in a rat brain complement DNA library using two probes A and B respectively of 350 and 380 bases. They were obtained by a polymerase chain reaction (PCR) carried out using the following primers
SEQ ID NO: 3 (Probe A, sense primer)
GACTGAGGAGCCCTTCCCTTTTCA
SEQ ID NO: 4 (Probe A, antisense primer)
GCCTTAGGATAGAAGTCATAGCCA
SEQ ID N 5 (Probe B, sense primer)
CCCTTTGTAGGAAAGGTTGTGCC
SEQ ID NO: 6 (Probe B, antisense primer)
GAATACGCAATAATCGGGAGGATAC
Sequencing indicated that the first clone is the rodent homologue of human tripeptidylpeptidase II (TPP II). In contrast, in the second, the sequence differs in the 5 'part (from nucleotide 293) which was not known to date.

 The invention therefore also relates to the nucleotide sequence (SEQ ID No. 1 and SEQ ID No. 2) coding for the new isolated protein.

 The sequence comprises a hydrophobic segment of twenty amino acids indicating the existence of a transmembrane segment. Thus, this new protein, although probably derived from the same gene as the previous one by an alternative splicing process, is presented as a serine ectopeptidase.

 Also, the present invention relates to a method of screening drugs by measuring the activity of membrane or homologous tripeptidyl peptidase II enzyme, using a model substrate of this enzyme.

 By homologous enzyme is meant both tripeptidylpeptidases that would be genetically modified as non-membrane tripeptidylpeptidases, especially soluble, such as the one above, isolated by Barlow R.M. et al.

The inventors have been able to develop inhibitors of the degradation of endogenous neuropeptides, in particular
CCK, by measuring the activity of said enzyme in the purified state but also more simply in the native state on membrane preparations from tissues using a TPP II model substrate.

Also, the subject of the present invention is also chemical compounds corresponding to the following general formula (I)

où R6 représente H, F, CH3 ou OCH2Ph,

in which
R1 represents a hydrogen or a linear or branched C1-C4 alkyl group
R2 represents a hydrogen or an alkyl group
C1-C2
- n = 0 or 1 and m = 0 or 1 with n different from m
- R3 represents a divalent radical consisting of an alkyl chain - (CH2) p - where p represents an integer ranging from 1 to 4 (inclusive ends), -CH2-CH (cis.F) -, - CH2S-, -CH2 -CH (CH2Ph) -, - (CH2) 2-CH (CONH2) -, a pattern

where R6 is H, F, CH3 or OCH2Ph,

R4 represents a hydrogen, an n.C6H13 group, CH20H, CH2-NHn.C4H9,
amide CO-NH-R5 wherein R5 represents hydrogen or a linear or branched C1-C6 alkyl group, - (CH2) 3-SCH3, -CH2Ph, -CH2C6H11,

 or carbonyl CO-R7 where R7 is n.C5H11 or NC5H10.

 Throughout the description, "Ph" refers to the optionally substituted phenyl radical.

 The compounds of formula (I) have several centers of asymmetry and the invention therefore covers the racemic mixtures as well as the different stereoisomers and their mixtures in any proportions.

 According to a first preferred variant, the subject of the invention is compounds of formula (I) as defined above in which n and m are indifferently equal to O or 1 with the proviso that n is different from m.

 According to a first aspect, the invention relates more particularly to a first group of these compounds in which R3 represents the bivalent radical - (CH2) 2-, which corresponds to p = 2 (proline skeleton).

 Compounds of this type are in particular the compounds given in examples hereinafter, references under the internal codes UCL 1330-J, UCL 1371-H, UCL 1381-J, UCL 1382-J, UCL 1391-J, UCL 1393-F , UCL 1394-F, UCL 1327-J, UCL 1331-J, UCL 1353-H, UCL 1387-J, UCL 1416-H (prepared according to route 1); UCL 1324-J (lane 2); UCL 1372-F3 (lane 5); UCL 1389-J (lane 6); UCL 1419-H (lane 7); UCL 1420-H (lane 8); UCL 1424-A (lane 9).

 Among this first group of compounds, those in which R 1 represents CH 2 CH 3, (CH 2) 2 CH 3, (CH 2) 3 CH 3 CH (CH 3) 2, CH 2 CH (CH 3) 2 and CH (CH 3) CH 2 CH 3, R 2 represents a hydrogen, R 4 represents the group amide CO-NH-R5 and R5 represents hydrogen are known.

According to another aspect, the invention relates more particularly to a second group of these compounds in which
R3 represents the cisfluoroethylene-CH2-CH (cis.F) motif (substituted proline skeleton), for example the compound referenced under the UCL 1395-F internal code (channel 3) described below.

 According to another aspect, the invention relates to a third group of these compounds in which R 3 represents the benzylethylene -CH 2 -CH (CH 2 Ph) - (substituted proline skeleton), for example the compound UCL 1433-H (lane 4) described above. .

 According to yet another aspect of the invention, the subject of the invention is a fourth group of compounds of this first variant in which R 3 represents the -SCH 2 - (thiaproline skeleton) motif, for example the UCL 1374-F compound (route 1 ) described below.

que le t t e té t raliydro i soqu i no 1 i ne dont les composés UCL 1421-H et UCL 1485-H (voie 3) sont des exemples décrits ci-après.According to yet another aspect of this first variant, the invention relates to a fifth group of these compounds in which R3 represents half

The following are examples of Examples: UCL 1421-H and UCL 1485-H (Route 3).

 According to other aspects of this first variant, the invention relates more particularly to compounds in which R 3 represents the divalent radical - (CH 2) 4 corresponding to p = 4 (perhydroazepine backbone) such as the compound UCL 1482-H (route 11) the radical - (CH 2) 3 corresponding to p = 3 (pipecol ine squelte) such as the UCL 1398-F compound (lane 4), the -CH 2 - radical corresponding to p = 1 (azetidine skeleton) such as the UCL compound 1373-F (lane 1).

(squelette indoline) dont des exemples sont les composés UCL 1397-H, UCL 1413-H, UCL 1414-H, UCL 1415-H, UCL 1423-H (voie 3) ; UCL 1446-A2 (voie 105 ; UCL 1486-H, UCL 1534-H, UCL 1535-A, UCL 1546-H (voie 12) décrits ci-après.According to a second particularly preferred variant, the subject of the invention is compounds of formula (I) in which n = 0 and m = 1 and in which R3 represents the unit

(indoline skeleton), examples of which are UCL 1397-H, UCL 1413-H, UCL 1414-H, UCL 1415-H, UCL 1423-H (channel 3); UCL 1446-A2 (Lane 105, UCL 1486-H, UCL 1534-H, UCL 1535-A, UCL 1546-H (Lane 12) described hereinafter.

(squelette perhydroindoline)
tels que le composé UCL 144?-H (voie 3) donné en exemple ci-après.According to another aspect of this second variant, the invention relates to compounds in which R3 represents the unit

(perhydroindoline skeleton)
such as the compound UCL 144 -H (lane 3) exemplified hereinafter.

(squelette isoindoline) @els que le composé UCL 1466-H (voie 4) décrit ci-après.Finally, according to a third variant, the subject of the invention is the compounds of formula (I) in which n = 1 and m = 0 and in which R3 represents the unit

(Isoindoline backbone) such as UCL compound 1466-H (lane 4) described hereinafter.

 According to another aspect of this third variant, the invention relates to the compounds in which R 3 represents the radical - (CH 2) 2 -CH (CO-NH 2) - (nipecotic skeleton) such as, for example, the compound UCL 1378-H ( 3) given below.

 Among these compounds, the radical R4 preferably represents the amide group CO-NH-R5. Compounds in which R4 is CO-R7 carbonyl are also preferred.

According to other variants, the radical R4 may represent a hydrogen or one of the groups CH20H, CH2-NH (n.C4H9).

The compounds of formulas (I) wherein R2 is hydrogen are also particularly preferred. In other cases, R1 and R2 each represent a methyl group or
R1 represents hydrogen and R2 represents an ethyl group.

 The inhibitory effect of these compounds was evaluated by measuring membrane TPP II activity and expressed by their apparent dissociation constant Ki as described below.

The preferred compounds according to the invention are the following compounds
2 (S) -aminobutyryl-L-prolinamide (UCL 1338-J)
L-valyl-L-proline n.hexylamide (UCL 1330-J)
1- (2 (S) -aminobutyryl) -L-proline 3- (methylthio) propylamide (UCL 1382-J)
1- (2 <5) -aminobutyryl) -L-proline n.pentylamide (UCL 1391
J)
1- (2 (S) -aminobutyryl) -L-proline n.butylamide (UCL 1371
H)
1- (2 (S) -aminobutyryl) -L-proline [2 (S) -methyl] butylamide (UCL 1381-J)
1- (2 (S) -aminobutyryl) -L-proline n -propylamide (UCL 1393
F)
1- (2 (S) -aminobutyryl) -L-proline iso.butylamide (UCL)
1394-F)
L-valyl-L-proline n-butylamide (UCL 1324-J)
- L-alanyl-L-prolyl-L-borovaline pinanediol ester (UCL
1372-F3)
L-Alanyl-L-prolyl-difluoro-L-borovaline borohydrate (UCL 1389-J)
1- (2 (S) -aminobutyryl) - (4 (S) -fluoro) -L-proline n-butylamide (UCL 1395-F)
1- (2 (S) -aminobutyryl) - (4 (S) -benzyl) -L-proline n-butylamide (UCL 1433-H)
2- (2 (S) -aminobutyryl) -1,2,3,4-tetrahydro-3 (S) -isoquinolinecarboxylic acid n-butylamide (UCL 1421-H)
1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid n-butylamide (UCL 1397-H)
1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid n -propylamide (UCL 1413-H)
1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid methylamide (UCL 1414-H)
1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid ethylamide (UCL 1415-H)
1- (2 (S) -aminobutyryl) -2 (R / S) - (5-methoxy) indoline carboxylic acid n-butylamide (UCL 1486-H)
1- (2 (S) -aminobutyryl) -2 (R / S) - (6-methoxy) indoline carboxylic acid n-butylamide (UCL 1546-H)
1- (2 (S) -aminobutyryl) -2 (R 1 S) - (5-fluoro) indoline carboxylic acid n-butylamide (UCL 1534-H)
1- (2 (S) -aminobutyryl) -2 (R / S) - (5-benzyloxy) indoline carboxylic acid n-butylamide (UCL 1535-H)
1- (2 (S) -aminobutyryl) -2 (S) -t (3aS, 7aS) -perhydro] -indolinecarboxylic acid n-butylamide (UCL 1442-H)
2- (2 (S) -aminobutyryl) -1 (R / S) -isoindoline carboxylic acid n-butylamide (UCL 1466-H) and their corresponding salts or hydrates.

 These compounds are preferred inhibitors according to the invention and have a Ki constant vis-à-vis the tripeptidyl peptidase enzyme according to the invention, less than or equal to 1 μM.

The following compounds are also preferred
1- (2 (S) -aminobutyryl) -2-hexylpyrrolidinamide (UCL 1419
H)
1- (2 (S) -aminobutyryl) -2 - ((1-oxo) hexyl] pyrrolidinamide (UCL 1420-H)
1- (2-Amino-2-methylpropionyl) -L-proline n-butylamide (UCL 1424-A)
1- (2 (S) -aminobutyryl) -2 (R / S) -perhydroazethane carboxylic acid n-butylamide (UCL 1482-H)
1- (2 (S) -aminobutyryl) -2 (S) -indolinylmethyl-N-butylamine (UCL 1446-A2) and their corresponding salts or hydrates.

Other very particularly preferred compounds are the compounds of formula (I) having an indoline backbone, namely the following compounds:
1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid n-butylamide (UCL 1397-H)
1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid n -propylamide (UCL 1413-H)
1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid ethylamide (UCL 1415-H)
1- (2 (S) -aminobutyryl) -2 (R / S) - (5-methoxy) indoline carboxylic acid n-butylamide (UCL 1486-H)
1- (2 (S) -aminobutyryl) -2 (R / S) - (5-fluoro) indoline carboxylic acid n-butylamide (UCL 1534-H) and their corresponding salts or hydrates.

 These compounds are particularly preferred inhibitors having a Ki constant not exceeding 0.02 μM.

 The subject of the present invention is also the processes for preparing the compounds of general formula (I) described above.

Also, the invention relates more particularly to a process (route 1) for the preparation of a compound of general formula (I) given above in which
R1 represents a linear or branched C1-C4 alkyl group
R2 represents a hydrogen
n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents - (CH2) 2-, -CH2S-, -CH2-; and
R4 represents CO-NH-R5, CO-R7
where R5 represents hydrogen or a linear or branched C1-C6 alkyl group, - (CH2) 3-S-CH3, -CH2Ph and R7 represents
NC5H10, CH2C6H11
characterized in that it comprises
i) the formation of a compound of formula (III)

wherein R1 and R2 have the meanings given above and X represents a protecting group, from a compound of formula (II)

esterified on its acid function by a group Y and in which X, R1 and R2 have the meanings given above, by reaction with L-proline
ii) amidation of the acid function of the compound (III) with the appropriate amine R5NH2 or R7H
where Rs and R7 have the meanings given above, to form the derivative (IV)

protected on its primary amine function by group X
iii) removing the X group of the derivative (IV), to obtain the desired compound (I).

The group Y preferably represents the motif

 Step i) is carried out in the presence of triethylamine and water.

 Furthermore, in step ii), the mixed anhydride of the acid (III) and isobutyl chloroformate is formed, which is then reacted in situ with the amine R5NH2.

 In the compounds prepared according to this route 1, R is preferably an ethyl or isopropyl group.

The invention also relates to a process (route 2) for the preparation of a compound of general formula (I) given above in which
R1 represents a linear or branched C1-C4 alkyl group
R2 represents a hydrogen or a methyl group
- n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents a group - (CH2) 2-; and
R4 represents a CO-NH-R5 group where R5 represents a hydrogen or a linear or branched C1-C6 alkyl group;
characterized by comprising i) forming a compound of formula (IV)

wherein R1, R2 and Rs have the meanings given above and X represents a protecting group, from a compound of formula (II)

esterified to its acid function using a group Y and in which R1, R2 and X have the meanings given above, by reaction with a prolineamide of formula (V)

in which R5 has the meaning given above
ii) removal of the X protecting group of the compound (IV), to obtain the desired compound (I).

Prolineamide (V) can be prepared by reacting the
L-proline

 protected on its amine function by a protecting group X and esterified on its acid function using a group Y, with the appropriate amine R5NH2.

The group Y preferably represents the motif

 To form the compound (IV), the prolineamide thus obtained is reacted with the compound (II) in the presence of water and triethylamine in a solvent such as tetrahydrofuran or dioxane.

 In the compounds prepared according to this route 2, R 1 is preferably ethyl or isopropyl and R 5 is preferably hydrogen or n-butyl.

The invention also relates to a process (route 3) for the preparation of a compound of general formula (I) given above in which
R1 represents a linear or branched C1-C4 alkyl group;
R2 represents a hydrogen
n = 0 and m = 1 and R3 represents a group -CH2-CH (cis.F)

R 4 represents a group -CH 2 OH, CO-NH-R 5 where R 5 represents a hydrogen or a linear or branched C 1 -C 6 alkyl group;
characterized in that it comprises
i) the preparation of a compound of formula (X)

avec un composé de formule (IX)

in which R 1, R 2, R 3 and R 4 have the meanings given above and X represents a protecting group, by reaction of a compound of formula (VIII)

with a compound of formula (IX)

in which R1, R2, R4 and X have the meanings given above
ii) removing the X group of the compound (X), to form the desired compound (I).

According to this route 3, the compound (VIII) is prepared by amidation of the acid function of a compound (VI)

protected on its amine function by a protecting group X and in which R3 has the meaning given above, with the appropriate amine R5NH2, to form the derivative (VII)

 in which R3, R4 and X have the meanings given above, then by elimination of the protective group X.

For this, the mixed anhydride of the acid (VI) and of isobutyl chloroformate is advantageously formed in the presence of
N-ethylmorpholine in tetrahydrofuran, which is then reacted in situ with the appropriate amine R5NH2. These conditions may also be those of step i) except that the compound (VII) is reacted in the place of the amine R5NH2.

 In the compounds prepared according to this route 3, R 1 is preferably ethyl and R 5 is preferably hydrogen, methyl, ethyl, n-propyl or n-butyl.

The invention also relates to a process (route 4) for the preparation of a compound of general formula (I) given above in which
R1 represents a linear or branched C1-C4 alkyl group
R2 represents a hydrogen
n = 1 and m = 0 and R3 represents a group -CH (CH2Ph)
CH2-, - (CH2) 3- or

R4 represents a CO-NH-R5 amide group where R5 represents a hydrogen or a linear or branched C1-C6 alkyl group;
characterized in that it comprises i) obtaining a compound (XII)

avec le composé de formule (XI)

protected on its primary amine function by a protective group X and in which R1, R2 and R3 have the meanings given above, by reaction of the compound of formula (IX)

with the compound of formula (XI)

in which R1, R2 and R3 have the meanings given above and X represents a protective group
ii) hydrolyzing the ester function of the compound (XII) thus obtained to form the compound of formula (XIII)

in which R1, R2, R3 and X have the meanings given above
iii) amidation of the acid function of the compound (XIII) using the appropriate amine R5NH2 to form the derivative of formula (XIV)

wherein R1, R2, R3, R5 and X have the meanings given above
iv) removing the X protecting group of the compound (XIV) to form the desired compound (I).

 The hydrolysis step ii) is advantageously carried out in the presence of sodium hydroxide in methanol.

 In step iii), the mixed anhydride of the acid (XIII) and isobutyl chloroformate is formed, which is then reacted in situ with the appropriate amine RsNH2.

 In the compounds prepared according to this route 4, R 1 is preferably ethyl and R 5 is preferably hydrogen or n-butyl.

caractérisé en ce qu'il comprend
i) l'amidation de la fonction acide du composé de formule (XIX)

The invention also relates to a process (route 5) for the preparation of a compound of general formula (I) given above, in which
- R1 represents a methyl group
R2 represents a hydrogen
- n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents the group - (CH2) 2-; and
- R4 represents a amide group CO-NH-R5 where Rs represents the unit

characterized in that it comprises
i) amidation of the acid function of the compound of formula (XIX)

pour former le dérivé (XX)

protected on its primary amine function by a protective group X and in which R1 and R2 have the meanings given above, using the amine (XVIII)

to form the derivative (XX)

in which R1, R2, R5 and X have the meanings given above
ii) removing the X protecting group of the compound (XX) to obtain the desired compound (I).

 Step i) is advantageously carried out using isobutyl chloroformate to form the mixed anhydride of the acid XIX, in the presence of N-ethylmorpholine in tetrahydrofuran.

 Step ii) can be carried out using trifluoroacetic acid in methylene chloride.

The amine (XVIII) is prepared
i) by reaction of isopropylboronic acid with (+) - pinanediol in ether, to obtain the derivative (XV)

ii) by reacting the compound (XV) with butyllithium and methylene chloride and then with zinc chloride ZnCl 2 in tetrahydrofuran, at low temperature, to form the derivative (XVI)

iii) substitution of the chlorine of the compound (XVI) with butyllithium and hexamethyldisilazane in tetrahydrofuran at low temperature, to form the compound (XVII)

 iv) releasing the amine function of the compound (XVII) to obtain the desired compound (XVIII).

 Isopropylboronic acid is obtained by adding triethylborate to isopropylmagnesium chloride in ether at low temperature.

 The above step iv) is advantageously carried out with trifluoroacetic acid in pentane.

The invention also relates to a process (route 6) for the preparation of a compound of general formula (I) given above in which
- R1 represents a methyl group
R2 represents a hydrogen
- n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents a group - (CH2) 2-; and
R4 represents a CO-NH-R5 amide group where R5 represents (CH3) 2 -CH-BF2
characterized in that it comprises
i) the elimination of the pinane unit of the compound of formula (I) wherein R 1, R 2, n, m R 3 and R 4 have the meanings given above and Rs represents

by action of boron trichloride in methylene chloride followed by hydrolysis, to obtain the derivative (XXI)

 ii) reacting the compound (XXI) with hydrofluoric acid to form the desired compound (I).

avec le composé de formule (IX)

The invention also relates to a process (route 7) for the preparation of a compound of general formula (I) given above in which
R1 represents a linear or branched C1-C4 alkyl group
R2 represents a hydrogen
- n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents a group - (CH2) 2-; and
- R4 represents the group n.C6H13
characterized in that it comprises
1) the reaction of a compound (XXV)

with the compound of formula (IX)

wherein R1 and R2 have the meanings given above and X represents a protecting group, to form the derivative of formula (XXVI)

in which R1, R2 and X have the meanings given above
ii) hydrogenating the double bond of the compound (XXVI) thus obtained to form the compound of formula (XXVII)

wherein R1, R2 and X have the aforementioned meanings;
iii) removing the X protecting group of the compound (XXVII) to obtain the desired compound (I).

Compound (XXV) is prepared from Lprolinal (XXIII)

protected on its heterocyclic amine function by a protecting group X, by reaction with triphenylpentylphosphonium bromide in the presence of butyllithium in toluene, to form the derivative (XXIV)

 and then removing the X protecting group of the compound (XXIV) thus obtained, for example using trifluoroacetic acid.

 L-proline (XXIII) can be prepared by reacting the L-proline methyl ester (XXII), protected on its amino function by a protecting group X, with diisobutylaluminum hydride in toluene at low temperature.

 The methyl ester of L-proline (XXII) can, for its part, be obtained by the action of thionyl chloride on Lproline in methanol.

 In the above-mentioned step i), the reaction of the protected 2 (S) -aminobutyric acid (IX) is carried out, for example, with isobutyl chloroformate to form its mixed anhydride in situ, in the presence of N- ethylmorpholine in tetrahydrofuran.

 The hydrogenation of step ii) can be carried out in methanol in the presence of Pd / C.

 The deprotection of step iii) can be obtained by the action of trifluoroacetic acid, then sodium hydroxide and then oxalic acid.

 In the compounds prepared according to this route 7, R preferably represents an ethyl group.

The invention also relates to a process (route 8) for the preparation of a compound of formula (I) given above, in which
R1 represents a linear or branched C1-C4 alkyl group
R2 represents a hydrogen
- n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents a group - (CH2) 2-; and
- R4 represents a carbonyl group CO-R7 where R7 represents the group n.C5H11
characterized in that it comprises
i) obtaining a compound of formula (XXX)

avec le composé de formule (IX)

wherein R1 and R2 have the meanings given above and X is a protecting group, by reaction of the compound (XXIX)

with the compound of formula (IX)

in which R1, R2 and X have the meanings given above
ii) removing the X protecting group of the obtained compound (XXX), to form the desired compound (I).

 In step i), the mixed anhydride of protected 2 (S) -aminobutyric acid (IX) and isobutyl chloroformate is formed and then reacted in situ with compound XXIX).

The compound (XXIX) can be obtained by hydrogenation of the corresponding hexagon (XXVIII)

 L-proline, obtained by the action of pentyllithium in ether, on L-proline protected on its amine function by an X group. The hydrogenation is advantageously carried out in the presence of Pd / C in methanol.

avec l'acide de formule (IX)

The invention also relates to a process (route 9) for the preparation of a compound of general formula (I) given above, in which
- R1 and R2 each represent a methyl group
- n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents a group - (CH2) 2-; and
R4 represents a CO-NH-R5 amide group where Rs represents a linear or branched C1-C6 alkyl group;
characterized in that it comprises
i) the reaction of the appropriate L-prolineamide of formula (V)

with the acid of formula (IX)

in which R1, R2 and R5 have the meanings given above and X is a protecting group, to form the derivative of formula (IV)

wherein R1, R2, R5 and X have the aforementioned meanings
ii) removing the protective group X of the compound (IV) to obtain the compound (I) sought.

 Step i) is advantageously carried out in the presence of dicyclohexylcarbodiimide in methylene chloride.

 L-prolineamide (V) can be obtained by forming the mixed anhydride of protected L-proline on its heterocyclic amine function and isobutyl chloroformate, in the presence of N-ethylmorpholine in tetrahydrofuran, which is then react with the appropriate amine R5NH2, which leads to the compound (V) desired after deprotection, for example by trifluoroacetic acid.

 In the compounds prepared according to this route 9, R 5 is preferably n-butyl.

- R4 représente le groupe CH2-NH(n.C4H9) caractérisé en ce qu'il comprend i) la formation d'un composé de formule (XXXII)

à partir de l'amide de formule (XXXI) correspondant

The invention also relates to a process (route 10) for the preparation of a compound of general formula (I) given above, in which
- R1 represents a hydrogen
- R2 is different from hydrogen
- n = 0 and m = 1 and R3 represents a pattern

- R4 represents the group CH2-NH (n.C4H9) characterized in that it comprises i) the formation of a compound of formula (XXXII)

from the corresponding amide of formula (XXXI)

in which X represents a protecting group
ii) protecting the secondary amine function of the compound (XXXII) thus formed by a group X ', to obtain the compound (XXXIII)

iii) selective deprotection of the heterocyclic amine by removal of the X group of the compound (XXXIII), to form the compound (XXXIV)

iv) the reaction of the compound (XXXIV) with the compound of formula (IX)

wherein R1, R2 and X have the above meanings to form a compound of formula (XXXV)

in which R1, R2 X and X 'have the meanings indicated above
v) removing the X and X 'protecting groups of the compound (XXXV) to form the desired compound (I).

 Step i) is carried out using diborane in tetrahydrofuran at low temperature.

 Step iv) is advantageously carried out using dicyclohexylcarbodiimide and hydroxybenzotiazole in a dimethylformamide / methylene chloride mixture.

 In the compounds prepared according to this route, R 2 is preferably ethyl.

à partir du composé de formule (IX)

The invention relates, moreover, to a process (route 11) for the preparation of a compound of general formula (I) given above, in which
R1 represents a linear or branched C1-C4 alkyl group
R2 represents a hydrogen
- n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents a group - (CH2) 4-; and
R4 represents a CO-NH-R5 amide group where Rs represents a linear or branched C1-C6 alkyl group;
characterized in that it comprises
i) formation of the compound of formula (XXXVI)

from the compound of formula (IX)

wherein R1 and R2 have the meanings given above and X is a protecting group, using 3-hydroxy-1,2,3-benzotriazine-4 (3H) -one in the presence of dicyclohexylcarbodiimide (DCC) in tetrahydrofuran
ii) reaction of the compound (XXXVI) with 2- (R / S) -perhydroazepine acid, to form the derivative (XXXVII)

wherein R1, R2 and X have the aforementioned meanings
iii) amidation of the acid function of the compound (XXXVII) thus obtained, using the appropriate amine R5NH2, to form the derivative of formula (XXXVIII)

wherein R1, R2 and X have the aforementioned meanings
iv) removing the X protecting group of the compound (XXXVIII) to obtain the desired compound (I).

 Step ii) is advantageously carried out in the presence of triethylamine in dimethylformamide.

 In step iii), the mixed anhydride of the acid (XXXVII) and isobutyl chloroformate is formed, which is then reacted with the appropriate amine R5NH2.

 In the compounds prepared according to this route 11, R 1 is preferably ethyl and R 5 is preferably n-butyl.

Finally, the invention relates to a process (route 12) for the preparation of a compound of general formula (I) given above, in which
R1 represents a linear or branched C1-C4 alkyl group
R2 represents a hydrogen
- n = 0 and m = 1 and R3 represents the motive

à partir de l'ester de formule (XXXXIII)

where R6 is OCH3, OCH2Ph, F; and
R4 represents a CO-NH-R5 amide group in which R5 represents a linear or branched C1-C6 alkyl group;
characterized in that it comprises
i) formation of the amide of formula (XXXXIV)

from the ester of formula (XXXXIII)

in which R6 has the meaning given above, by reaction with the appropriate amine R5NH2
ii) the reaction of the compound (XXXXIV) with the compound of formula (IX)

wherein R1 and R2 have the aforementioned meanings and
X represents a protecting group, to form the compound of formula (XXXXV)

in which R1, R2, Rg, R6 and X have the meanings given above
iii) removing the X group to form the desired compound (I).

 Step ii) is advantageously carried out in the presence of bis (2-oxo-3-oxazolodinyl) phosphinic acid chloride and triethylamine in methylene chloride.

pour former le composé de formule (XXXX)

As for the methyl ester (XXXXIII), it can be obtained
i) by the action of sodium nitrite in the presence of hydrochloric acid, on the compound of formula (XXXIX)

to form the compound of formula (XXXX)

in which R6 has the meaning indicated above
ii) by adding ethyl 2-methylacetoacetate to the compound (XXXX) thus obtained, in the presence of sodium nitrite in ethanol, to form the compound of formula (XXXXI)

in which R6 has the above meaning
iii) by cyclization in acidic medium, to form the ethyl ester (XXXXII)

 iv) by exchange from the ethyl ester (XXXXII) thus obtained, in the presence of magnesium in ethanol.

 The ethyl ester undergoes ester exchange and reduction in the presence of magnesium and methanol to yield the methyl ester (XXXXIII).

 In the compounds prepared according to this route 12, R 1 is preferably ethyl and R 5 is preferably n-butyl.

 In all the methods described above, the formation of the acid anhydrides can be carried out by conventional methods, advantageously using isobutyl chloroformate in the presence of N-ethylmorpholine in tetrahydrofuran.

 Furthermore, according to the invention, X or X 'represents a conventional protecting group capable of protecting the desired function in a given molecule without altering the other functions thereof, such as in particular benzyl, benzyloxycarbonyl or tert-butoxycarbonyl groups. These groups are introduced by conventional methods well known to those skilled in the art.

 It is the same for the deprotection steps which are also carried out by methods known per se such as in particular acid hydrolysis or catalytic hydrogenation especially in the presence of Pd / C.

 As for the group Y, it is a radical capable of esterifying an acid function without modifying the other functions of the molecule, and constituting a good leaving group in order to be easily removed in a subsequent step. It is preferably the succinimide ester.

 The work of the applicant has shown that the compounds of formula (I) have numerous therapeutic properties and in particular in the field of the treatment of disorders or conditions in which the inactivation of endogenous neuropeptides is involved, such as in particular the disorders or conditions inactivation of CCK.

 Also, the subject of the present invention is also the use of a compound of formula (I) as a medicament for treating, in humans or in animals, in particular disorders of food intake, of the mood, cognitive or motor disorders, including anorexia, schizophrenia, Parkinson's disease and depression as well as gastrointestinal transit disorders such as irritable bowel syndrome.

 The present invention also relates to the use of a compound of formula (I) in the preparation of a medicament for the treatment, in humans or animals, of conditions or disorders caused by the physiological degradation of neuropeptides endogenous in particular that of the CCK.

 The present invention particularly relates to the use of compounds of formula (I) in the preparation of a medicament for the treatment of disorders or conditions as mentioned above.

 Compounds of formula (I) useful as medicaments can be administered in a physiologically acceptable carrier.

 Also the subject of the present invention is pharmaceutical compositions comprising a therapeutically effective amount of a compound of formula (I) in combination with a physiologically acceptable carrier.

 The present invention further relates to the use of the compounds of formula (I) as inhibitors of the inactivation of endogenous neuropeptides and in particular that of CCK.

The inventors have found that, in general, among the compounds of formulas (I), those having a chirality L, L (that is to say S, S according to the nomenclature Ingold
Kahn-Prelog) are the most active. In the examples given below, the configuration of the other optically active carbons is also indicated for the compounds of formulas (I) having more than two centers of asymmetry.

 In all the abovementioned uses, it is preferred to choose the compounds according to the invention having a Ki constant of less than or equal to 1 μM, such as those exemplified and, in a most preferred manner, the compounds of formula (I) having a indoline skeleton.

According to the invention, all the aforementioned uses also include that of the compounds of formulas (I) already known, as indicated above, that is to say the compounds of formula (I) in which R1 represents a CH2CH3 group, (CH2 ) 2CH3, (CH2) 3CH3, CH (CH3) 2, CH2CH (CH3) 2 or CH (CH3) CH2CH3,
R2 represents hydrogen, R3 represents the divalent radical - (CH2) 2-> R4 represents a CO-NH-R5 amide group and Rs represents hydrogen.

 Other advantages and characteristics of the present invention will appear on reading the experimental results which follow, in particular of a method for isolating and characterizing the tripeptidyl peptidase according to the invention, as well as examples of preparation given by way of illustration and non-limiting.

ISOLATION AND CHARACTERIZATION OF CCK-INACTIVATED ENZYME
The inventors used brain membranes
(rat cerebral cortex) as original material and unsulfated CCK-8 peptides (Asp-Tyr-Met-Gly-Trp-Met-Asp-Phe
NH2) and CCK-5 (Gly-Trp-Met-Asp-Phe-NH2) as substrates, the characteristic products of the reaction (CCK-5 and Gly-Trp-Met) being measured fluorimetrically after isolation by high performance liquid chromatography ( HPLC) according to Camus et al., (Neurosci., 1989, 29: 595).

 The successive HPLC purification steps involved are described in the following Table I which also indicates the purification factors obtained using CCK-8 as a substrate.

TABLE I
Retention time
Steps or Molarity of Factor
[eluent] eluent at the peak purification
business
Membranes i
Membrane solubilization 0.4
(detergent: 1%)
Ion exchange (DEAE 5PW) 300 mM 15
[NaCl 0-300 mM]
Hydroxyapatite (Bio-Gel HPHT) 220 mM 45
[phosphate 0-300 mM]
Gel filtration (Protein Pak 300su) 45 min 3,500
[50 mM phosphate]
Hydrophobic column (Phenyl-5 PVV) 0 8.571
[(NH4) 2 S04 1M-O]
From the second stage of purification, the chromatography profile has only one peak of enzymatic activity which is superimposed on the peak obtained by measuring the hydrolysis of CCK-5, indicating that a single enzyme is responsible for both. cuts of the CCK-8 molecule.

 After the last purification step, sodium dodecyl sulfate (SDS) gel electrophoresis indicates a single band irreversibly labeled with DFP-3H at an apparent mass of 135 kDa.

The study of the specificity of the purified enzyme on a series of model substrates has shown that it behaves like an aminotripeptidylpeptidase capable, in particular, of hydrolyzing the Ala-Ala-Phe-p.nitroanilide or Ala fragments.
Ala-Phe-amidomethylcoumarin (A-AP-Amc) releasing nitroaniline or aminomethylcoumarin.

EXAMPLES OF PREPARATION OF COMPOUNDS OF FORMULA (I)
For the spectral data given in all the examples below, the following abbreviations have the following meanings:
s = singlet, m = multiplet, d = doublet, dd = doublet of doublets, t = triplet, dt = doublet of triplets, q = quadruplet, tq = triplet of quadruplets and tt = triplet of triplets.

Example 1: Preparation of L-valyl-L-proline n.hexylamide (UCL 1330-J); lane 1
R1 = CH (CH3) 2; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-NH-R5; Rs = (CH2) 5CH3
a / Preparation of N-benzyloxycarbonyl-L-valine
10 g of L-valine (85 mmol) are dissolved in 4 M KOH (40 ml) and the mixture is cooled in ice. 21.9 g of benzyloxycarbonyl chloride (128 mmol) are added over 5 minutes and a white solid is formed. The mixture is stirred at 0 ° C. for one hour and then the temperature is allowed to rise and stirring is continued for another hour at room temperature. The reaction mixture is then diluted with 4M KOH (200 ml) and extracted with ether (2 x 100 ml). The aqueous phase is then brought to pH 1 with concentrated hydrochloric acid and then the ethyl acetate extract (3 x 200 ml). The extracts are combined and dried (Na2SO4) and the solvent is evaporated. 21.99 g of a clear oil are obtained.

 1 H-NMR (200 MHz, CDCl 3, ppm): 7.42-7.52 (1H, d, NH) 7.24-7.36 (5H, m, Ph) 5.00 (2H, s, Ph. CH2) 3.78-3.88 (1H, m, α-CH acid); 1.98-2.08 (1H, m, P-CH acid); 0.80-0.88 (6H, m, 2 x CH3).

b / Preparation of the succinimide ester of
N-benzyloxycarbonyl-L-valine
Dissolved in dimethoxyethane (100 ml) was 15.63 g (62 mmol) of the product obtained in a / and 7.16 g of N-hydroxysuccinimide (62 mmol) and the mixture was cooled in ice. 12.8 g of dicyclohexylcarbodiimide (62 mmol) are added with stirring and the mixture is refrigerated overnight. The white solid which forms is filtered off and the filtrate is concentrated to give 24.28 g of a white solid by crystallization from isopropanol. 13.20 g of the desired product are obtained. BP = 105-6 ° C. (Literature: 116-7 ° C.)
1 H-NMR: 7.35 (5H, s, ArCH); 5.27-5.41 (1H, d, NH); 5.11 (2H, s, PhCH 2); 4.58-4.71 (1H, m, -CH valine), 2.80 (4H, s, 2 x CH2), 2.19-2.41 (1H, m, F-CH valine); 99-1.07 (6H, m, 2 x CH3).

c / Preparation of N-benzyloxycarbonyl-L-valyl-L-proline
3.4 g of L-proline (29.9 mmol) are dissolved in water (100 ml) and 4.5 g of triethylamine (44.8 mmol) are added. Tetrahydrofuran (100 ml) was dissolved in 10 g (29.9 mmol) of the product obtained under cooling with ice. The proline solution is added in 40 minutes and the mixture is stirred for one hour at 0 ° C. and then at room temperature overnight. The solvent volume is reduced by about half, acidified (pH <1) with concentrated HCl and extracted with ethyl acetate (3 x 200 ml). The organic phases are combined and dried (Na 2 SO 4) and the solvent is evaporated. 11.95 g of an oil are obtained.

 TLC analysis (ethyl acetate / acetic acid 18) showed the presence of 4 components.

The product obtained is redissolved in ethyl acetate and extracted with 4M KOH (3 x 200 ml). The aqueous phases (pH <1) are acidified with concentrated HCl and extracted with ethyl acetate (3 x 200 ml). The combined organic phases are dried (Na 2 SO 4) and the solvent is evaporated. 8.09 g of an oil (77.7%) is obtained
HPLC (column: Lichrosorb rp select B, eluent CH3OH / H20 7/3 + 0.1% trifluoroacetic acid) indicated purity> 90%.

 1 H-NMR (200 MHz, CDCl 3, ppm): 7.25-7.30 (5H, m, Ph) 5.90-5.95 (1H, d, NH) 5.04-5.10 (2H, m.p. m, PhCH2); 4.50-4.57 (1H, m, -CH valine); 4.26-4.34 (1H, m, -CH proline); 3,633.79 (2H, m, CH 2 proline); 1.95-2.14 (4H, m, 2 x CH2proline) 0.87-1.00 (6H, m, 2 x CH3).

 Mass Spectrum (EI): 349 (M + + 1).

d / Preparation of N-benzyloxycarbonyl-L-valyl-L-proline hexylamide
Tetrahydrofuran (50 ml, dry, distilled) is dissolved in 2 g (5.7 mmol) of the product obtained in c / and is cooled in an ice / salt bath. 660 mg of
N-ethylmorpholine (5.7 mmol) and 780 mg of isobutyl chloroformate (5.7 mmol) and the mixture was kept cool for 30 minutes. 580 mg of n-hexylamine (5.7 mmol) are added, the temperature of the mixture is allowed to warm to room temperature and the mixture is left overnight. The mixture was dissolved in water (50 ml) and extracted with ethyl acetate (2 x 50 ml). The organic phases are combined and dried (Na 2 SO 4) and the solvent is evaporated. 2.95 g of a brown oil are obtained. Crystallization from an ethyl acetate / petroleum ether mixture gives 370 mg of a white powder. The filtrates are concentrated. Chromatography using ethyl acetate as eluent gives 1.80 g (73.2%) of the desired product.

 HPLC (column: Kromasil C-18, eluent: CH 3 OH / H 2 O 7/3 + 0.1% trifluoroacetic acid) indicated> 90% purity.

1 H-NMR (200 MHz, CDCl 3, ppm) 7.33 (5H, s, Ph); 5.455.50 (1H, d, NH); 5.07-5.08 (2H, d, PhCH2); 4.52-4.57 (1H, m, -CH valine); 4.28-4.36 (1H, m, -CH proline); 3.56-3.72 (2H, m, CH 2 proline); 3.11-3.21 (2H, m, α-CH 2 amide); 1.69-2.39 (5H, m, P-CH valine, 2 x CH 2 proline); 15-1.45 (8H, m, 4 x
CH2); 0.81-0.98 (9H, m, 3 x CH3).

e / Preparation of the title compound as oxalate
900 ml (208 mmol) of the product obtained in d) are dissolved in methanol (100 ml) and 300 mg of palladium on charcoal are added. The mixture is hydrogenated for 3 hours and then the catalyst is filtered off. After removal of the solvent, 602 mg (94.5%) of a pale oil are obtained.

 TLC analysis showed that no starting material remained.

 182 mg of oxalic acid (202 mmol) in 5 ml of ethanol are added and the oil is dissolved. After addition of ether (50 ml), a turbid solution is obtained which is refrigerated. 660 mg (84%) of the desired product is obtained as the oxalate salt.

Mp 146-7 ° C
Elemental analysis: C16H31N302.C2H2O4
Found: C = 55.99%; H, 8.75%; N = 10.63%;
Theoretical: C = 55.80%; H, 8.58%; N = 10.84%
1 H-NMR (200 MHz, DMSO, ppm): 7.85-7.91 (1H, t, NH) 4.25-4.32 (1H, q, -H proline); 3.93-3.96 (1H, d, αH valine) 3.43-3.72 (2H, m, CH 2 proline); 2.95-3.07 (2H, m, aCH 2 amide) 1.66-2.15 (5H, m, P-CH valine, 2 x CH 2 proline); 1.25-1.39 (8H, m, 4 x CH 2); 0.8-1.2 (6H, m, 3 x CH3).

 IR (cm-1): 3400, 3333 (N-H), 2700-3200 (OH), 1683, 1639 (C = O), 1195 (CONH).

HPLC (column: Kromasil C-18, eluent: CH3OH / H2O 1/1 + 0.1% trifluoroacetic acid): 100%
Mass Spectrum (FAB): 298 (M + + 1).

Example 2: Preparation of 1- (2 (S) -aminobutyryl) -Lproline 3- (methylthio) propylamide (UCL 1382-J); lane 1
R1 = CH2CH3; R2 = H; nn = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2- R4 = CO-NH-R5; R5 = (CH2) 3SCH3
a / Preparation of N- (tert.butoxycarbonyl) 2 (S) -aminobutyric acid
Dissolve in a mixture of dioxane (155 ml), water (78 ml) and 1N NaOH (78 ml), 8 g of 2 (S) -aminobutyric acid (77.6 mmol). The solution is cooled in an ice / salt bath and 18.62 g of di-tert.butyl dicarbonate (85.4 mmol) are added. The solution was allowed to warm to room temperature and stirred for one hour. The mixture was then concentrated in vacuo to a volume of about 90 ml and then acidified with KHSO4 solution.

The resulting solution was extracted with methylene chloride (2 x 200 ml), the combined extracts were dried and evaporated to give an oil. Extensive drying under vacuum gives 14.20 g of a white waxy solid (91%). PF = 65-66C
bJ Preparation of succinimide ester of N- (tert.butoxycarbonyl) -2 (S) -aminobutyric acid
In dry tetrahydrofuran, under nitrogen at 0 ° C., 14.2 g of acid (70 mmol) obtained in a /. 14.4 g of dicyclohexylcarbodiimide (70 mmol) and 8.1 g of N-hydroxysuccinimide (70 mmol) are added, and the mixture is stirred overnight. The precipitate formed is filtered off, rinsed once with ethyl acetate and the solvent was removed under vacuum resulting in 18.00 g of an oil (85%). The product is used in subsequent steps without further purification.

 1 H-NMR (CDCl3, ppm): 4.6 (1H, m, aH); 2.6 (4H, s, CH 2 CH 2 succinimide); 1.5-1.7 (2H, m, CH 2 aminobutyryl); 1.2 (9H, s, (CH3) 3); 0.9 (3H, t, CH3aminobutyryl).

c / Preparation of 1- (N- (tert.butoxycarbonyl) 2 (S) -aminobutyryl) -L-proline
6.933 g of ester (23.1 mmol) obtained in b) are dissolved in tetrahydrofuran (150 ml) and the mixture is cooled to 0 ° C. A solution of 3.99 g of L-proline (34.degree. 7 mmol) and 3.51 g of triethylamine (34.7 mmol) in water (50 ml). Stirring is maintained for 48 hours at room temperature. The tetrahydrofuran is removed under vacuum and replaced with water and the pH is brought to about 2 with diluted KHSO4. The acidic solution was extracted with methylene chloride, the extracts were dried and evaporated to give an oil (64% yield).

1 H-NMR (CDCl3, ppm): 4.3 (1H, m, H), 4.2 (1H, m, H) 3.6-3.8 (2H, m, CH2Nproline); 1.9-2.1 (4H,
CH2CH2CH2Nproline); 1.6-1.8 (2H, m, CH 2 aminobutyryl); 1.5 (9H, s, (CH3) 3); 0.9 (3H, t, CH3aminobutyryl).

d / Preparation of 1- (N- (tert.butoxyearbonyl) -2 (S) -aminobutyryl) -L-proline 3- (methylthio) propylamide
Tetrahydrofuran (100 ml), under nitrogen, was dissolved in 2.47 g (8.23 mmol) of the product obtained under c /, while cooling in an ice / salt bath. 0.948 g of N-ethylmorpholine (8.23 mmol) and 1.125 g of isobutyl chloroformate (8.23 mmol) are added. After 30 minutes, 0.864 g of 3- (methylthio) propylamine (8.23 mmol) are added. Stirring is maintained overnight. The suspension is poured into methylene chloride (250 ml), washed with 10% citric acid (3 x 100 ml) and then with 5% NaHCO3 (3 x 100 ml), dried and evaporated. which leads to an oil (60% yield).

1 H-NMR (CDCl 3, ppm): 4.4 (1H, m, aH); 4.3 (1H, m, alpha) 3.5-3.8 (2H, m, CH 2 Nproline); 3.2 (2H, m, CH 2 N); 2.2 (2H, m,
CH2CH2Nproline); 2.0 (3H, s, CH3S); 1.6-1.8 (8H, m,
CH2CH2CH2Nproline, CH2CH2SCH3, CHaminobutyryl); 1.5 (9H, s, (CH3) 3); 0.9 (3H, t, CH3aminobutyryl).

e / Preparation of the title compound as oxalate salt
Chilled trifluoroacetic acid (100 ml) was dissolved in 5 g (12.9 mmol) of the product obtained in d / under cooling in an ice / salt bath and the mixture was stirred for 30 minutes. The acid is removed under vacuum, which leads to a clear oil. This oil is treated with 5% NaHCO3 and extracted with methylene chloride. The organic phase is dried and evaporated, which leads to the free base (clear oil). The resulting oil is dissolved in a small amount of ethanol and oxalic acid (1.1 equivalents) is added. After addition of ether, white crystals of the desired product are obtained as the oxalate salt (25% yield). This product is sufficiently pure without recrystallization being necessary.

PF = 120-121 ° C
1 H-NMR (CD30D, ppm): 4.5 (1H, m, alphaH); 4.3 (1H, m, aH) 3.6 (2H, m, CH2Nproline); 2.5 (2H, m, CH 2 NH); 2.1 (3H, s,
SCH3) 1 1.75-2.0 (10H, m, SCH2CH2CH2CH2CH2Nproline, CH2 aminobutyryl); 1.0 (3H, t, CH3aminobutyryl).

HPLC (column: Lichrosorb rp select B: eluent
CH30H / H2O 30/70): 99.99% (Tr = 8.66 minutes).

Elemental analysis: C13H25N3O2S.C2H2O4.0,5H2O
Found: C, 47.17, 9w; H = 7.05%; N = 10.67%
Calculated: C, 46.62%; H, 7.30%; N = 10.87%
Example 3 Preparation of 1- (2 (S) -aminobutyryl)
L-proline n.pentylamide (UCL 1391-J); lane 1
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) -; R4 = CO-NH-R5; Rs = (CH2) 4CH3
a / Preparation of N-benzyloxycarbonyl-2 (S) -aminobutyric acid
4 g of 4M KOH (100 ml) are dissolved in 10 g of 2 (S) -aminobutyric acid (97 mmol) and cooled to 0 ° C. 24.7 g of benzyl chloroformate (145 mmol) are added. and stirring the mixture allowing the temperature of the mixture to rise to room temperature for 2 hours 30 minutes. The mixture is then diluted with water (100 ml) and extracted with ether (2 x 200 ml). The aqueous phase is acidified to pH <1 with concentrated HCl and extracted with ethyl acetate (3 x 200 ml). The ethyl acetate fractions are combined, dried (Na 2 SO 4) and the solvent removed. 20.4 g of a white solid are obtained (yield 89%).

1 H-NMR (ppm): 7.32 (5H, s, ArCH); 5.22-5.28 (1H, d,
NH); 5.10 (2H, s, Ph-CH 2); 4.02-4.44 (1H, m, a-CH); 1.582.02 (2H, m, CH 2); 0.9-1.2 (3H, t, CH 3).

b / Preparation of the succinimidic ester of the acid
N-benzyloxycarbonyl-2 (S) -aminobutyric
Dissolve in dimethoxyethane (160 ml), 20 g of acid (84 mmol) obtained in a / and 9.7 g of N-hydroxysuccinimide (20 mmol) and cool the solution in ice. 17.3 g of dicyclohexylcarbodiimide (84 mmol) are added portionwise with cooling and a white precipitate forms rapidly. The mixture is cooled overnight, filtered and the solvent removed. 30.69 g of a viscous oil are obtained.

Crystallization from isopropanol gives 5.74 g of a white solid.

1 H-NMR (ppm): 7.31 (5H, s, ArCH); 5.3-5.6 (1H, d, NH) 5.1 (2H, s, Ph-CH 2); 4.8-4.9 (1H, m, a-CH); 2.9 (4H, s, 2 x
CH2ester); 1.80-2.05 (2H, m, CH 2 aminobutyryl); 0.95-1.25 (3H, m, CH3aminobutyryl).

c / Preparation of 1- (N-benzyloxycarbonyl-2 (S) -amino butyryl) -L-proline
5 g of the ester (14.95 mmol) obtained in b / and dissolved in tetrahydrofuran (50 ml) are dissolved in ice.

1.75 g of L-proline (14.95 mmol), 2.25 g of triethylamine (22.4 mmol) and water (50 ml) are then added. The mixture is stirred and cooled for one hour and then warmed to room temperature overnight. The solvent content is reduced to about half its volume, acidified and extracted with ethyl acetate (3 x 100 ml). The combined ethyl acetate fractions (Na 2 SO 4) are dried and the solvent is removed. 5.73 g of a thick oil are obtained.

1 H-NMR (ppm): 7.38 (5H, s, ArCH); 5.9-6.0 (1H, d, NH) 5.08 (2H, s, Ph-CH 2); 4.42-4.54 (2H, m, 2 x a-CH); 3.52-3.85 (2H, m, CH 2 -proline); 1.52-2.30 (6H, m, 2 x CH2proline,
CH 2aminobutyryl); 0.85-1.0 (3H, m, CH 3).

d / Preparation of 1- (N-benzyloxycarbonyl-2 (S) -amino butyryl) -L-proline n.pentylamide
The 2 g (5.98 mmol) of the product obtained at 0 ° C. was dissolved in dry, distilled tetrahydrofuran (50 ml) and cooled in an ice / salt bath. 0.813 g of isobutyl chloroformate (5.98 mmol) and 0.687 g of N-ethylmorpholine (5.98 mmol) were added with stirring and the solution became cloudy. Stirring and cooling were continued for 30 minutes and then 0.52 g of n-pentylamine (5.98 mmol) was added. The mixture is stirred at room temperature overnight and then diluted with water (50 ml) and extracted with ethyl acetate (2 x 50 ml). The combined organic extracts are dried (Na2504) and the solvent is removed. 2.54 g of an oil are obtained. HPLC and TLC analyzes showed that there was still starting acid. The resulting oil is redissolved in ethyl acetate (100 ml), washed with 4M KOH (50 ml) and water and dried (Na2504) and the solvent removed. 2.03 g of the desired product is obtained (yield 84%).

1 H-NMR (ppm): 7.33 (5H, s, ArCH); 5.07-5.08 (2H, m,
CH2Ph); 3.96-4.07 (2H, m, a-CH); 3.48-3.9 (2H, m, CH 2 proline); 1.1-2.5 (12H, m, 6 x CH2); 0.75-1.0 (6H, m, 2 x
CH3).

e / Preparation of the title compound as oxalate
1 g (2.48 mmol) of the product obtained in d) was dissolved in methanol (100 ml) and 200 mg of Pd / C (10%) were added. This mixture is hydrogenated under pressure for 3 hours in a Parr apparatus. The catalyst is removed by filtration and the solvent is removed and 223 mg of oxalic acid (2.48 mmol) are added. Crystallization from an ethanollether mixture gives 290 mg of desired product (yield 32.5%).

Mp = 140-141 ° C
HPLC (eluent: CH30H 30% / H2O / 0.1% TFA): 99.3%
Elemental analysis: C14H27N302.0.9C2H204.0.2C2H50H.O.5H2O
Found: C, 52.63%; H, 8.44%; NN = 11.24%
Calculated: C = 52.78%; H = 8.48% N = 11.40%
Mass Spectrum (FAB): 270 (M + + 1)
1 H-NMR (DMSO-d6, ppm): 7.64-7.69 (1H, m, NH); 4.24-4.30 (1H, m, a-CH); 4.03-4.09 (1H, m, a-CH); 3.30-3.74 (2H, m,
CH2proline); 2.90-3.10 (-NH-CH 2 -); 1.52-2.10 (6H, m, 2 x
CH2proline, CH2 butyl chain); 0.72-1.44 (12H, m, 2 x CH 3, 3 x CH 2 pentyl chain).

Example 4 Preparation of 1- (2 (S) -aminobutyryl)
L-Proline N-Butylamide (UCL 1371-H); lane 1
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-NH-R5; R5 = (CH2) 3CH3
a / Preparation of 1- (N- (tert.butoxycarbonyl) -2 (S) -aminobutyryl) -L-proline n-butylamide
2.42 g of 1- (N- (tert.butoxycarbonyl) -2 (S) -aminobutyryl) -L-prolate (8.05 mmol), prepared as in Example 2, are dissolved in dry tetrahydrofuran (15 ml). c /. The solution is cooled in an ice / salt bath and then 0.927 g of
N-ethylmorpholine (8.05 mmol) and 1.10 g of isobutyl chloroformate (8.05 mmol). The solution is stirred for 45 minutes and 0.59 g (8.05 mmol) of n-butylamine are added.

After stirring overnight, methylene chloride (200 ml) was added and the mixture was washed with 10% citric acid (2 x 100 ml) and then with 5% NaHCO3 (2 x 100 ml).

The organic phase is dried (Na 2 SO 4) and evaporated under vacuum.

We get a syrup. Deprotection is carried out without characterization.

b / Preparation of the title compound as oxalate
The product obtained with stirring in an ice / salt bath is treated with cold trifluoroacetic acid (TFA) (28 ml) for 30 minutes. The TFA is removed under vacuum. The resulting oil is treated with 5% cold NaHCO 3 until the pH is about 10 and then extracted with methylene chloride. The organic phase is dried (Na 2 SO 4) and evaporated. The oil is taken up in a small volume of ethanol and treated with oxalic acid. After addition of ether, white crystals of the desired product are obtained as the oxalate salt.

This compound is sufficiently pure without recrystallization (50%).

Mp 165-167 ° C
1 H-NMR (CD30D, ppm): 4.4 (1H, m, aH); 4.2 (1H, m, aH) 3.4-3.7 (2H, m, CH2N proline); 3.0-3.2 (2H, m, CH2NC4Hg) 1.6-2.2 (4H, m, NCH2CH2CH2 proline); 1.2-1.5 (4H, m, CH 2 aminobutyryl, NCH 2 CH 2 Cl 4 H 9); 1.1 (3H, t, CH 3 aminobutyryl) 0.9 (3H, t, n-butyl CH 3).

HPLC (column: Lichrosorb rp select B, eluent
CH3OH / H2O 55/45 + 0.1 * TFA): 99.99% (Tr: 4.82 minutes).

Elemental analysis: Cl3H25N302.C2H204.1,3H20
Found: C, 49.15 *; H, 7.62%; N = 11.06%
Theoretical: C = 48.85%; H, 8.09%; N = 11.39%
Example 5 Preparation of 1- (2 (S) -aminobutyryl)
L-proline E2 (S) -methylbutylamide (UCL 1381-J); lane 1
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different than m; R3 = - (CH2) 2- R4 = CO-NH-R5; R5 - (S) CH2CH (CH3) CH2CH3
Preparation of 1- (N-benzyloxycarbonyl-2 (S) -amino-butyryl) -L-proline
It is treated with a solution of 4.138 g of N-benzyloxycarbonyl-2 (S) -aminobutyric acid succinimide ester (12.4 mmol) prepared as in Example 3b / in dimethoxyethane (25 ml), a solution of 2.139 g of L-proline (18.6 mmol) and 1.56 g of NaHCO 3 (18.6 mmol) in water (20 ml). After 5 hours, 12.4 ml of water are added, the resulting mixture is acidified to pH = 2 with concentrated HCl and extracted with ethyl acetate. The extracts are dried and evaporated to yield a syrup (48% yield).

1 H-NMR (CDCl 3, ppm): 7.2 (5H, m, H-Aryl); 5.2 (1H, m, alphaH); 5.0 (3H, m, CH 2 benzyloxycarbonyl, aH); 4.0-4.1 (2H, m,
CH2N); 1.8-2.0 (4H, m, CH 2 CH 2 CH 2 N); 1.1-1.2 (2H, m,
CH 2aminobutyryl); 0.9 (3H, t); 3.8 (2H, m, CH 2 Aryl); 3.5 (2H, m, CH 2cycle); 3.2 (2H, m); 2.0 (2H, m); 0.9 (3H, t,
CH3aminobutyryle).

b / Preparation of 1- (N-benzyloxycarbonyl-2 (S) -aminobutyryl) -L-proline [2 (S) -methyl) butylamide
Tetrahydrofuran (100 ml), 3.8 g (11.4 mmol) of the product obtained in (a) was dissolved and the mixture was cooled under nitrogen in an ice / salt bath. 1.31 g of
N-ethyl-morpholine (11.4 mmol) and then 1.487 g of isobutyl chloroformate (11.4 mmol). After 30 minutes, 0.99 g of 2 (S) -methylbutylamide (11.4 mmol) are added and stirring is continued overnight. The suspension is poured into methylene chloride (250 ml), washed with 10% citric acid (3 x 100 ml) then with 5% NaHCO3 (3 x 100 ml), dried and evaporated. An oil is obtained (yield 50%).

1 H-NMR (CDCl 3, ppm): 7.2 (5H, m, H-Aryl) 5.0 (2H, d,
CH 2 benzyloxycarbonyl); 4.3-4.5 (2H, m, 2H); 3.2-3.6 (2H, m,
CH2N) 2 2.9-3.1 (2H, m, CH2Nproline) 1.0-2.1 (18H, m,
CH2CH2CH2N, CH3CH2aminobutyryl, NCH2CH (CH3) CH2CH3).

c / Preparation of the title compound as oxalate
1.96 g of the product obtained in (b) are dissolved in methanol and 0.24 g of palladium-on-charcoal (10%, wet) is added. This suspension is hydrogenated in a Parr apparatus under 40 psi for 4 hours. The catalyst is removed by filtration and the solvent removed in vacuo. The resulting oil is dissolved in a little ethanol and oxalic acid (1.1 equivalents) is added. After addition of ether, the desired product is obtained as an oxalate salt (yield 45%).

Mp = 135-136 ° C
1 H-NMR (CD30D, ppm): 4.4 (1H, m, aH); 4.25 (1H, m, aH) 3.65 (2H, m, CH2Nproline); 3.0 (2H, m, CH 2 NH); 2.0 (5H, m, CH 2 CH 2 Nproline, CH 3 CH 2 CH (CH 3) CH 2 N), 1.5 (2H, m,
CH 2aminobutyryl); 1.1 (3H, t, CH3aminobutyryl); 0.9 (6H, m,
CH3CH2CH (CH3) CH2 N).

 HPLC (Lichrosorb column, select B, eluent CH3OH / H2O 15/85 + 0.1% TFA): 99.99% (Tr = 4.63 minutes).

Elemental analysis: C 14 H 27 N 3 O 2 .C 2 H 2 O 4 .5H 2 O
Found: C, 52.04%; H, 7.82%; N = 11.27%
Calculated: C, 52.16%; H, 8.21%; N = 11.41%
Example 6: Preparation of 1- (2 (S) -aminobutyryl) -Lproline n.propylamide (UCL 1393-F); lane 1
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-NH-R5; R5 = (CH2) 2CH3
a / Preparation of 1- (N- (tert.butoxyzarbonyl) -2 (S) -aminobutyryl) -L-proline n -propylamide
Tetrahydrofuran (40 ml) was dissolved in 2 μl of 1- (N- (tert.butoxycarbonyl) -2 (S) aminobutyryl) -L-proline (6.6 mmol) prepared as in Example 2 c /. 0.86 ml of iso-ethyl chloroformate (6.6 mmol) and 0.85 ml of N-methylmorpholine (6.6 mmol) are then added to this solution at -10 ° C. and under nitrogen. After stirring at -10 ° C. for 20 minutes, 2.75 ml of n-propylamine (33.0 mmol) are added.

The mixture is stirred at 0 ° C. for one hour and then at room temperature overnight. The solvent is evaporated in vacuo and a white semi-solid material is obtained. It is dissolved in ethyl acetate (150 ml), washed with 5% citric acid (30 ml), saturated sodium bicarbonate solution (30 ml) and brine (20 ml). ). After drying, Na 2 SO 4 is filtered off and the solvent is evaporated. An oil is obtained which is purified by liquid chromatography on a column (eluent: ethyl acetate) and 2.06 g of a white solid are obtained (yield 90%).

 1 H-NMR (CDCl 3, ppm): 6.86 (1H, broad 6, NH); 5.22 (1H, d, NH); 4.56 (1H, dd, &alpha;-CHaminobutyryl); 4.37 (1H, m, α -CH proline); 3.54 (2H, m, N-CH2proline); 3.13 (2H, m, N-CH 2 propyl); 2.39-1.31 (8H, m, 4 x CH 2); 1.24 (9H, s, (CH 3) 3 -) 0.91 (3H, t, CH 3 propyl); 0.86 (3H, t, CH 3 aminobutyryl).

Mass Spectrum (FAB): 342 (MH +)
b. Preparation of the title compound as trifluoroacetate
The product obtained in (a) was dissolved in dichloromethane (5 ml), 2.0 g (5.9 mmol) and trifluoroacetic acid (5 ml) was added to this cooled solution. The reaction mixture is stirred for 2 hours in an ice bath.

The acid is evaporated and precipitation is obtained with ethyl ether and dried overnight at 30 ° C. under 0.1 mmHg, 1.34 g of desired product (yield 59%).

PF = 46-51 C (open capillary)
HPLC: 99.8%
1 H-NMR (D 2 O / TSP, ppm): 4.41 (1H, t,? - chaminobutyryl) 4.33 (1H, t, -CHproline) 3.71 (2H, m, N-CH 2 -proline); 3.17
(2H, dt, N-CH 2 amide) 2.21 (2H, dt, CH 2 -proline); 1.86 to 2.03
(4H, m, 2 x CH 2); 1.54 (2H, tq, CH 2 amide); 1.04 (3H, t, CH 3 aminobutyryl); 0.89 (3H, t, CH3 amide).

Mass Spectrum (FAB): 242 (MH +)
Elemental analysis: C12H23N302.1,3CF3COOH
Found: C, 44.75%; H, 6.13%; N = 10.59%
Calculated: C = 45.01%; H, 6.29%; N = 10.79%
Example 7: Preparation of 1- (2 (S) -aminobutyryl) -Lproline iso-butylamide (UCL 1394-F); lane 1
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-NH-R5 = = CH2CH (CH3) 2
a / Preparation of 1- (N- (tert.butoxycarbonyl) -2 (S) -aminobutyryl) -L-proline iso.butylamide
The procedure is as in Example 6a / using iso-butylamine instead of n-propylamine. 2.21 g of an oil are obtained (yield 94%).

1 H-NMR (CDCl 3, ppm): 6.93 (1H, brs, NH); 5.24 (1H, brs, NH); 4.56 (1H, dd, = -CHaminobutyryl); 4.36 (1H, tt, α-CHproline); 3.53 (2H, m, N-CH 2 -proline); 3.00 (2H, m, NCH 2); 2.39-1.58 (7H, m, CH and CH2); 1.37 (9H, s, (CH 3) 3-) 0.94
(3H, t, CH 3); 0.84 (6H, d, 2 x CH3).

Mass Spectrum (FAB): 356 (MH +)
b / Preparation of the title compound in trifluoroacetate form
The procedure is as in Example 6b / from the product prepared in a / and 0.62 g of desired product is obtained (yield 29%) by precipitation with ethyl ether and drying at room temperature under vacuum (0.degree. , 1 mmHg) overnight.

PF = 58-61 C (open capillary)
HPLC = 100%
1 H-NMR (D 2 O / TSP, ppm): 4.43 (1H, t,? - chaminobutyryl) 4.33 (1H, t, ', -CHproline); 3.70 (2H, m, N-CH 2 -proline); 3.04 (2H, dd, N-CH 2 amide); 2.23 (2H, m, CH 2 primary); 1.96 (4H, m,
CH2); 1.78 (1H, m, CH); 1.04 (3H, t, CH3aminobutyryl); 0.89 (6H, d, 2 x CH3).

Mass Spectrum (FAB): 256 (MH +)
IR (KBr pellet, cm-1): 3298 and 3089 (NH), 2962 (CH), 1662 (C = O).

Elemental analysis: C13H25N3O2.1,2CF3COOH
Found: C, 47.15%; H, 6.75%; N = 10.72%
Calculated: C, 47.16%; H, 6.73%; N = 10.71
Example 8: Preparation of L-valyl-L-proline piperidineamide (UCL 1327-J); lane 1
R1 = CH (CH3) 2; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-R7; R7 =

 The procedure is as in Example 1.

Mp = 56-58 ° C
1 H-NMR (400 MHz, CD 3 OD, ppm): 3.89 (1H, broad, α-CH) 3.39 (1H, brs, α-CH); 3.35 (4H, m, N- (CH2) 2-); 3.24 (2H, t, N-CH 2); 2.09 (2H, brs, proline CH2); 1.82 (2H, brs, CH2proline); 1.61 (1H, m, CH (CH3) 2); 1.47 (2H, tt, -CH 2 -); 1.36 (2H, broad t, -CH 2 -); 0.95 (3H, d, CH 3); 0.86 (3H, d, CH3).

 IR (nujol paste, cm-1): 3445 and 3419 (N-H amine), 1744 (C = O acid), 1648 (C = O tertiary amide).

Elemental analysis: C13H23N302. 2.1 (COOH) 2.0,3H20
Found: C, 48.55%; H = 7.00%; N = 8.71%
Calculated: C, 48.46%; H, 6.74%; N = 8.83%
Example 9: Preparation of L-valyl-L-proline n.butylamide (UCL 1324-J); lane 2
R1 = CH (CH3) 2; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) -; R4 = CO-NH-R5; R5 = (CH2) 3CH3
a / Preparation of N- (tert.butoxycarbonyl) -L-proline n.butylamide
5.0 g of succinimide ester of N- (tert.butoxycarbonyl) -L-proline (0.016 mmol) sold by NOVABIOCHEM are dissolved in tetrahydrofuran (50 ml). 1.533 ml of n-butylamine (0.015 mmol) are then added thereto over 10 minutes and the mixture is stirred for one hour. The solvent is removed and the resulting crude material is purified by chromatography on silica using as eluent ethyl acetate. 4.3 g of an oil are obtained (yield 100%).

 1 H-NMR (200 MHz, CDCl 3, ppm): 6.3 (1H, brs, NH); 4.25 (1H, m, methine) 3.6-3.1 (4H, m, methylene); 2.9 (2H, m, methylene, 1.95 (9H, s, methyl), 1.6-1.2 (6H, m, methylene).

b / Preparation of L-proline n.butylamide, trifluoroacetate
4.22 g of the product obtained in (0.014 mmol) and 15 ml of trifluoroacetic acid are stirred at room temperature for one hour. The excess acid is evaporated under vacuum and dried under vacuum for 3 hours. 4.5 g of desired product are obtained (yield 100%).

 1 H-NMR (400 MHz, CDCl 3, ppm): 9.3 (1H, brs, NH) 7.65 (1H, brs, NH); 4.7 (1H, m, methine); 3.5 (2H, m, methylene); 3.25 (2H, m, methylene); 2.1 (4H, m, methylene) 1.5 (2H, m, methylene); 1.3 (2H, m, methylene); 0.9 (3H, t, methyl).

c / Preparation of N- (tert.butoxycarbonyl) -L-valyl
L-proline n.butylamide
Tetrahydrofuran (3.times.77 g) of succinimide ester of N- (tert.butoxycarbonyl) -L-vallne (0.0117 mol) sold by NOVABIOCHEM is dissolved in tetrahydrofuran (30 ml). A solution of 3.35 g of the product obtained in b / (0.0117 mol) in 20 ml of tetrahydrofuran is added with stirring at 25 ° C. 3.0 ml of triethylamine are then added to neutralize the trifluoroacetate salt and the mixture is stirred for 20 hours The solvent is evaporated and the resulting crude material is purified by chromatography on silica using a mixture of petroleum ether and ethyl acetate 1/1 as eluent.

1.8 g of desired product (yield 41%) are obtained in the form of a colorless oil.

1 H-NMR (200 MHz, CDCl 3, ppm): 6.9 (1H, brs, NH); 5.2
(1H, d, NH); 4.5 (1H, dd, methine); 4.25 (1H, dd, methine) 3.6 (2H, m, methylene); 3.2 (2H, q, methylene); 1.1 (5H, m, methine and methylene); 1.4 (9H, s, CH3tert.butyl); 1.3 (4H, m, methylene); 0.9 (6H, dd, methyl); 0.8 (3H, t, methyl).

d / Preparation of L-valyl-L-proline n.butylamide, trifluoroacetate
1.5 g of the obtained product (c / 4.8 mmol) and 6 ml of trifluoroacetic acid are stirred at 25 ° C. for 45 minutes, the excess acid is evaporated and the resulting crude material is purified. by chromatography on silica using as eluent a mixture of ether / ethyl acetate / acetic acid (1/1 / 0.1) then ethyl acetate / methanol / acetic acid (1/1 / 0.1). further purification by preparative HPLC under the following conditions: Lichrosorb column select B, eluent: 60/40 ethanol / trifluoroacetic acid 0.1% The product is obtained as an oil.

e / Preparation of the title compound as oxalate
20 g aqueous Na2CO3 (5 ml), 1.7 g of the product obtained at d (4.4 mmol) are dissolved, stirred for 10 minutes and extracted with ethyl acetate (3x). 15 ml). The organic extracts are dried (Na 2 SO 4) and concentrated to give an oil (0.734 g). 1 g of oxalic acid is dissolved in 3 ml of ethanol and this solution is added to the oil obtained above, the mixture is heated in a water bath for 10 minutes and then cooled. By addition of ethyl ether, a white precipitate is obtained which is filtered and washed with ethyl ether. It is recrystallized from methanol / ethyl acetate / ether (1/1/3), which gives 0.680 g of white crystals (42% yield).

Mp = 165-166 ° C
TLC (ethyl acetate / methanol / acetic acid 6/2/2)
Rf = 0.25
IR (KBr, cm-1): 3432 (NH), 3072 (+ NH3), 2958 (CH), 1718 (C = O), 1641 (C = O), 1554 (NH).

 1 H-NMR (200 MHz, DMSO-d 6, ppm): 7.9 (1H, t, NH); 7.3 (2H, brs, NH 2); 4.25 (1H, t, methine) @ 3.95 (1H, d, methine); 3.65 (1H, m, CH 2 proline); 3.45 (1H, m, CH 2 proline); 3.0 (2H, m, methylene); 2.2-1.5 (5H, m, methine and methylene); 1.3 (4H, m, methylene); 0.9 (6H, dd, methyl) 0.8 (3H, t, methyl).

 Mass spectrum: 269 (M +) (2%).

Elemental analysis: C16H29N306.0.2 (COOH) 2
Found: C, 52.12%; HH = 8.01%; N = 11.07%
Calculated: C, 52.19%; H = 7.85%; N = 11.13%
Example 10 Preparation of L-Alanyl-L-Prolyl
L-borovaline pinanediol ester (UCL 1372-F3); lane 5
R1 = CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-NH-R5 R5 =

a / Preparation of isopropylboronic acid
A solution of 149 g of triethylborate (1 mole) in 300 cm3 of ether and a 2M solution of 103 g of isopropylmagnesium chloride (1 mole) in 500 cm 3 of dichloromethane are added simultaneously under a nitrogen atmosphere and vigorously stirring. ether, in ethyl ether (500 cm3) stirred under nitrogen at -78 ° C, keeping the temperature below -70 ° C for 2 hours. The temperature of the reaction mixture is allowed to rise to room temperature and the stirring is continued for a further 24 hours. The mixture is then acidified with sulfuric acid diluted to 40% cold (250 cm 3) and stirred while maintaining the temperature. below 15 C. Stir for an additional 16 hours and further dilute with water (250 cm3) to remove the emulsion.

The ether phase is separated and the aqueous phase is extracted with ether (3 × 200 cm 3). The ethereal phases are combined and dried (magnesium sulphate) and then concentrated. The resulting solid is crystallized from water. 75 g (wet) of desired product is obtained which must be stored wet or in ether to avoid autooxidation.

 1 H-NMR (DMSO-d6), ppm): 3.4 (2H, brs, OH); 0.8 (7H, s, methine and methyl).

 [Isopropylboronic acid has been described by R. M. Washburn et al., Org. Syn., 1963, Coll. Flight. IV: 68-72].

b / Preparation of hexahydro-2 (1-methylethyl) -3a, 5.5 trimethyl- (3a5- (3aa, 4a, 6a, 7aa)] -4,6-methano-1,3,2-benzodioxaborol (or isopropylboronic acid pinanediol ester): compound
XV.

30 g of acid (0.34 mol), prepared at 40 g of (+) - pinanediol (0.23 mol) in 150 cm3 of ether, are stirred for 48 hours. The reaction mixture is diluted with 150 cm3 of ether and 100 cm3 of water and the ethereal phase is then separated off. The aqueous phase is extracted with ether (3 × 50 cm 3), the organic phases are combined, dried (magnesium sulfate) and concentrated. The resulting oil is purified by chromatography on silica eluting with a mixture of petroleum ether / ethyl acetate (9/1). The oil obtained is further purified by vacuum distillation and 28 g of the desired product are recovered with a boiling point of 64-68 ° C. under 0.3.
Torr (45% yield).

 1 H-NMR (CDCl 3, ppm): 4.25 (1H, dd, methine); 2.4-1.7 (6H, m, methylene and methine); 1.35 (3H, s, methyl); 1.25 (3H, s, methyl); 1.0 (7H, s, methyl and methine); 0.8 (3H, s, methyl).

 [This compound has been described by C. A. Kettner et al., J. Biol.

Chem., 1984, 259, 151061.

 c / Preparation of 2- (1-chloro-2-methylpropyl) -hexahydro-3a, 5,5-trimethyl- [3α5- [2 (1S), 3αa, 4α, 6α, 7α, -4,6-methanol; 1,3,2-b enzodioxaboroie: compound XVI.

 Stirred at -105 C under nitrogen, 200 cm3 of tetrahydrofuran and 10.6 g of dichloromethane (0.124 mole). 40 ml of 1.6M butyllithium in hexane (0.084 mol) are introduced by means of a syringe at -105 ° C. in 40 minutes. This mixture is stirred for 15 minutes and a mixture of 15 g of compound XV (0.0675 mol) obtained in b / in 60 cm3 of tetrahydrofuran is slowly introduced, with the aid of a syringe, while maintaining the temperature at The reaction mixture is stirred for 30 minutes and then 40.5 cm 3 of 1M zinc chloride in ethyl ether (0.0405) is introduced at -10 ° C. in a syringe, using a syringe. The temperature is allowed to rise to room temperature and stirred for a further 16 hours.The solvent is removed under reduced pressure and the resulting oil is taken up in 500 cm3 of ether.

The ether extract is washed with a saturated aqueous solution of ammonium chloride (100 cm3) and the aqueous phase is separated. It is extracted with ether (2 × 100 cm 3), the combined ethereal solutions are dried (magnesium sulfate) and concentrated.

The resulting oil is vacuum distilled and 26.9 g of desired compound XVI is obtained in the main fraction (88% yield) with a boiling point of 74-108 ° C at 0.2 Torr.

1 H-NMR (CDCl 3, ppm) 4.35 (1H, d, O-CH-pinanediol ester); 3.32 (1H, d, CHCl); 2.45-1.9 (7H, m, methylene methine ester of pinanediol); 1.35 (3H, s, methyl); 1.25
(3H, s, methyl); 1.0 (6H, dd, CH3isopropyl) 0.8 (3H, s, methyl).

d / Preparation of hexahydro-3a, 5,5-trimethyl-α- (1-methylethyl) -N, N-bis (trimethylsilyl) - [3a5- [2 (R), 3aa, 4a, 6a, 7aa] - 4,6-methano-1,3,2-benzodioxaborole-2-methaneamine: compound XVII
The mixture is stirred at -72 c under nitrogen, 24.3 cm3 of hexamethyldisiiazanne (0.115 mole) and 120 cm3 of tetrahydrofuran. 61 cm3 of 1.6M n.butyllithium in hexane (0.0974 mol) are gradually added thereto at -72 ° C. to -70 ° C. in 40 minutes. The temperature is allowed to rise to 0 ° C. for 3 hours and the reaction mixture is cooled again to -72 ° C. A mixture of 24 g of compound XVI (0.0886 mol) is introduced slowly over 30 minutes at -72 ° C. obtained in 100 cm 3 of tetrahydrofuran. The temperature is then raised to 20 ° C. and the mixture is stirred for 20 hours and then concentrated. The resulting oil is treated with ether (250 cm 3) and water (100 cm 3). The organic phase is separated off and the aqueous phase is extracted with ethyl ether (2 × 100 cm 3) and the ethereal solutions are then combined, dried (sodium sulfate) and concentrated. The resulting oil is vacuum distilled (removing the head) to give 32 g of compound XVII (91% yield) with a boiling point of 120-155 ° C. at 0.2 Torr.

 1 H-NMR (CDCl 3, ppm): 4.25 (1H, d, O-CH pinanediol ester); 2.4-1.7 (7H, m, methylene and methine); 1.35 (3H, s, methyl); 1.25 (3H, s, methyl); 1.15 (1H, d, methine) 0.9 (6H, dd, CH3isopropyl); 0.8 (3H, s, methyl).

e / Preparation of hexahydro-3a, 5,5-trimethyl-α- (1-methylethyl) - [3a5- [2 (R), 3aa, 4a, 6a, 7aa] -4,6-methano-1,3 2-benzo dioxaborole-2-methaneamine, trifluoroacetate (or borovaline trifluoroacetate pinanediol ester) compound XVIII
0 g (0.0258 mol) of the product obtained in d / in 10 cm3 of pentane are stirred at 0 ° C. under a nitrogen atmosphere. 0.584 cm3 of trifluoroacetic acid (0.0758 mol) is slowly introduced into it and this mixture is stirred for one hour at 0 ° C. After concentration under reduced pressure and then suctioning under high vacuum for 48 hours, compound XVIII is obtained with quantitative yield, in the form of a white solid.

 1 H-NMR (DMSO-d 6, ppm): 7.7 (2H, brs, NH 2); 4.3 (1H, d, O-CH); 2.8 (1H, m, a-CH); 2,4-1,8 (7H, m, methine and methyl); 1.35 (3H, s, methyl); 1.25 (3H, s, methyl); 1.0 (6H, dd, methyl); 0.8 (3H, s, methyl).

[a1D23 + 25 "(c 1.0, CHCl3)
f / Preparation of N- (tert.butoxycarbonyl) -L-alanyl-
L-proline: compound XIX
30 cm3 of 0.5M aqueous NaOH and 30 cm3 of dioxane, 3 g of L-alanyl-L-proline in the form of hydrate (0.0147 mol) are dissolved and the mixture is cooled to 0 ° C. with stirring.

0.53 g of di-tert.butyl dicarbonate (0.0162 mol) are introduced at 0 ° C. with the aid of a syringe. The reaction mixture is stirred at 0 ° C. for 30 minutes and then at 200 ° C. for one hour. 30 cm3 of ethyl acetate are added and the mixture is cooled to 0 ° C. It is then treated with dilute aqueous KHSO4 until pH = 2 and a precipitate forms. It is filtered, washed with ether and dried in vacuo. 3.17 g of desired product is obtained.

NMR (DMSO-d6, ppm): 6.9 (1H, d, NH); 4.2 (2H, m, - CH alanine and proline); 3.5 (2H, m, methylene); 2.1 (1H, m,
CH2proline); 1.9 (3H, m, CH 2 -proline); 1.3 (9H, s, methyl) 1.1 (3H, d, methyl).

[This compound has been described in particular by E. Wuensch et al.,
Int. J. Pept. Protein Res., 1988, 32, 3683.

g / Preparation of N- (tert.butoxycarbonyl) -L-alanyl
L-prolyl-L-borovaline pinanediol ester: compound XX
30 g of stirred tetrahydrofuran are dissolved under nitrogen, 3.17 g of the compound obtained in f / and cooled to -20 ° C.

2.27 g of N-ethylmorpholine (0.0190 mol) and then 1.59 g of isobutyl chloroformate (0.0116 mol) are introduced at -20 ° C. and a precipitate forms. The reaction mixture is stirred at -20 ° C. for 30 minutes and then a mixture of 4.05 g of compound XVIII is introduced in the form of trifluoroacetate (0.0110 mol), as obtained at e / in 20 cm3 of tetrahydrofuran. The mixture was stirred at -20 ° C for 30 minutes and then allowed to warm to 20 ° C and held there for 30 minutes. 30 cm3 of water are then added and the mixture is extracted with ethyl acetate (3 × 50 cm 3). The combined organic phases are dried and concentrated. The resulting oil is purified by chromatography on silica eluting with a chloroform / methanol (9/1) mixture. Drying the resulting oil in vacuo gives 3.32 g of the desired compound (yield 57%) of a white solid.

 1 H-NMR (CDCl 3, ppm): 7.0 (1H, d, NH); 5.4 (1H, d, NH) 4.65 (1H, d, a-CH); 4.45 (1H, t, a-CH); 4.30 (1H, d, O-CH) 3.5 (2H, m, methine); 3.1 (1H, t, α-CHborovaline); 2.5-1.5 (11H, m, methine and methylene proline and pinane); 1.4 (9H, s, methyl and tert.butyl); 1.35 (3H, s, methyl); 1.3 (3H, dd, methylalanine); 0.9 (6H, dd, methyl); 0.75 (3H, s, methyl).

h / Preparation of the title compound as trifluoroacetate
0.5 g of the compound obtained in g / (0.000962 mol) are dissolved in dichloromethane (2 cc) and the mixture is cooled under nitrogen. At 0 ° C. under nitrogen, 3.7 g of trifluoroacetic acid (0.0324 mol) are introduced and the mixture is stirred for 30 minutes. The excess acid is removed under reduced pressure and then under vacuum. The resulting oil is purified by preparative HPLC on a Kromasil column equipped with a detector at 215 nm eluting with 0.1% aqueous CF3COOH / 0.1% methanolic CF3COOH / 3/7 and an incolor solid is obtained which has a retention time of 9.7 minutes.

Mp = 124-125 ° C
IR (KBr, cm-1): 3475 (NH 2), 1671 (C = O amide), 1614 (C = O amide).

 1 H-NMR (CDCl 3, ppm): 7.9 (1H, s, NH); 4.5 (brs, NH2 and a-CH alanine); 4.2 (1H, d, B-O-H); 3.95 (1H, m, α-CH proline); 3.45 (2H, m, CH 2 proline); 2.4-1.6 (11H, m, methylene and methine); 1.4 (3H, d, CH3 alanine); 1.3 (3H, s, methyl); 1,2 (4H, s, methyl and methine); 0.8 (6H, dd, methyl); 0.7 (3H, s, methyl).

Elemental analysis: C22H38BN304.2CF3COOH
Found: C, 47.8%; H = 6.50%; N = 6.31%
Calculated: C = 48.2%; H, 6.23%; N = 6.49%
Example 11 Preparation of L-alanyl borohydrate
L-prolyl-difluoro-L-borovaline (UCL 1389-J); lane 6
R1 = CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-NH-R5; R5 =

 A solution of 0.20 g of 1M BCl 3 in CH 2 Cl 2 (0.000384 mol) is cooled to -50 ° C. A solution of 0.20 g of the compound obtained in Example 11 (0.000384 mol) in 1 cm 3 of CH 2 Cl 2 is added using a syringe under a nitrogen atmosphere and stirring. This reaction mixture is stirred for 15 minutes and stirring is continued at 0 ° C. for one hour.

1 cm3 of water and then 5 cm3 of ethyl acetate are added thereto. The aqueous phase is separated and evaporated to dryness at 20 ° C. The resulting white solid is dissolved in water (2 cc) and acetone (1 cc) and treated with 0.5M aqueous hydrofluoric acid (5 cm3) for 5 hours. 15 minutes. The solvent is removed at 20 ° C. under high vacuum, the residual solid is stirred in methanol and the insoluble residue is removed by filtration and the filtrate is evaporated to give a white solid which is crystallized from an acetone / acetone mixture. water / ether 0.55 g of the desired compound is obtained as borohydrate.

Mp = 207-208 ° C
IR (KBr, cm-1): 3439 (NH), 3190 (NH3), 1652 (C = O amide), 1629 (C = O amide), 727 (BF).

 1 H-NMR (D 2 O, ppm): 4.8 (1H, m, α-CH proline); 4.5 (1H, m, α-CH alanine); 3.8 (2H, m, CH 2 proline); 2.31 (1H, m, csCH valine); 2.0 (4H, m, CH 2 proline); 1.7 (1H, m, CH valine) 1.55 (3H, d, CH3 alanine); 0.88 (6H, dd, methyl).

Elemental analysis: C12H22BF2N3O2.0.95 H3B03
Found: C, 41.51%; H = 6.86; N = 11.72%
Calculated: C, 41.43%; H, 7.20%; N = 12.08%
Example 12: Preparation of 1- (2 (S) -aminobutyryl) -2hexyl pyrrolidineamide (UCL 1419-H); lane 7
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = n.C6H13
a / Preparation of the methyl ester of N- (tert.butoxycarbonyl) -L-proline: compound XXII
12.5 g of proline (0.108 mol) are dissolved in methanol (100 ml) and cooled to 5.degree. C. 8.75 ml of thionyl chloride (0.12 mol) are slowly added and refluxed. After the removal of the solvent, 20.6 g of a yellow oil are obtained and this oil is then dissolved in 200 ml of dimethylformamide and 21.4 g of triethylamine (0.2 mole) are added. The mixture is stirred for 30 minutes and then filtered and the filtrate is added in portions to 21.8 g of di-tert.-butyl dicarbonate (0.108 mol), the mixture is slowly degassed and then left to dry. After the removal of the solvent, an oily solid is obtained, which is chromatographed on silica gel, eluting with ethyl acetate to give 17.25 g of the desired ester.
(96.9% yield).

Mass spectrum: m / e = 229 (M +)
1 H-NMR (ppm): 4.0-4.18 (1H, m, NCHCO); 3.65 (3H, s, CH 3 O); 3.28-3.55 (2H, m, CH 2 cycle); 1.77-2.16 (4H, m, 2 x
CH2 cycle); 1.24-1.39 (9H, m, tert.C4HgO).

b / Preparation of N- (tert.butoxycarbonyl) -L-prolinal compound XXIII
1.5 g (0.065 mole) of the ester prepared in the reaction medium are dissolved under argon in dry toluene (300 ml) and cooled to -78 ° C.. M solution of diisobutylaluminum hydride in hexane
(0.159 moles). After 30 minutes at -60 ° C, methanol is added
(10 ml) and poured the reaction mixture into a solution of potassium sodium tartrate M (250 ml) and stirred at room temperature for 2 hours and allowed to stand overnight. The phases are separated and the aqueous phase is extracted with ether (2 × 200 ml). The combined organic phases are dried (Na 2 SO 4) and the solvent is removed. 14.71 g of a black oil are obtained.

Mass spectrum: m / e = 199 (M +)
1 H-NMR (ppm): 9.4-9.56 (1H, m, CHO); 3.76-4.20 (1H, m,
NCHCO); 3.22-3.66 (2H, m, CH 2 cycle); 1.58-2.16 (4H, m, 2 x
CH2 cycle); 1.30-1.52 (9H, m, tert.C4HgO).

c / Preparation of N- (trt.butoxycarbonyl) -2- (1-hexenyl) pyrrolidine: compound XXIV
21.89 g (0.053 mol) of pentyltriphenylphosphonium bromide (prepared according to the method of Jaenicke et al., Liebigs Annalen der Chemie, 1973: 1259) are suspended in argon in 260 ml of tetrahydrofuran (dry, distilled). and cooled to -78 ° C. 40 ml of a 1.6M butyllithium solution are added and the mixture becomes dark orange. 13 g (0.057 mol) of the compound prepared in solution in 30 ml of tetrahydrofuran (dry, distilled) are then added over 5 minutes. We
allow the mixture to warm to room temperature and
leave it overnight at this temperature. Then 250 ml of brine is added and a white precipitate is formed. The phases are separated and the aqueous phase is extracted with petroleum ether (2 x 250 ml). The combined organic phases are dried (Na 2 SO 4) and the solvent is removed. By a new addition of petroleum ether, a solid precipitates which is removed by filtration and the filtrate is concentrated. 3.5 g of a clear oil are obtained by column chromatography on silica gel, eluting with a mixture of ethyl acetate / petroleum ether 1/9.

Further chromatography was performed using ethyl acetate / petroleum (1:19) as eluent and 2.63 g of desired product was obtained.

Mass spectrum: m / e = 253 (M +)
1 H-NMR (400 MHz, ppm): 5.29-5.31 (2H, br m, -CH = CH-) 4.47 (1H, brs, N-CH-CH = CH-); 3.36-3.39 (2H, broad m, CH 2 cycle); 1.56-2.11 (6H, m, 2 x CH2 cycle, allylic CH2) 1.27-1.46 (14H, m, tert.C4HgO, 2 x CH2 chain); 0.86-0.90 (3H, m, CH 3).

d / Preparation of 2- (1-hexenyl) pyrrolidine compound XXV
20 ml of trifluoroacetic acid are cooled in an ice / salt bath and 2.3 g (9.09 mmol) of the compound prepared in c / are added. Stirring and cooling are continued for one hour, then the acid is removed and the residue is partitioned between 100 ml of a solution of sodium hydroxide and 100 ml of ether. The phases are separated and the aqueous phase is extracted with ether (2 x 100 ml). The fractions obtained are combined and dried (Na 2 SO 4) and the solvent is removed. 1.29 g of desired product are obtained (yield 92.8%).

Mass spectrum: m / e = 154 (M + + 1)
1 H-NMR (400 MHz, ppm): 5.37-5.55 (2H, m, -CH = CH-); 3,733.79 (1H, m, N-CH-CH = CH-); 2.81-2.89 (1H, m, -CH (H) -N ring) 2.61-2.67 (1H, m, -CH (H) -N ring) 1.95-2.12 ( 3H, m, allylic CH 2, CH (H) ring); 1.66-1.73 (1H, m, CH (H) ring) 1.37-1.56 (2H, m, CH 2 cycle); 1.20-1.34 (4H, m, 2 x CH 2) 0.84-0.92 (3H, m, CH 3).

Preparation of 1- (N- (tert.butoxycarbonyl) 2 (S) -aminobutyryl) -2- (1-hexenyl) pyrrolidinamide: Compound XXVI
N- (tert.butoxycarbonyl) -2 (S) -aminobutyric acid (795 mg, 3.92 mmol), prepared as in Example 2 (a), was dissolved in tetrahydrofuran (dry, distilled) and cooled to room temperature. C. 450 mg (3.92 mmol) of
N-ethylmorpholine and 3.92 mmol of isobutyl chloroformate and the mixture is stirred for one and a half hours. 600 mg (3.92 mmol) of the compound prepared in d) are then added and the mixture is left at room temperature overnight. The solvent is removed and the residue is dissolved in 100 ml of water and then extracted with ethyl acetate. The extracts are combined and dried (Na2SO4) and the solvent is removed. 1.62 g of a black oil are obtained which is purified by column chromatography on silica gel, eluting with ethyl acetate / acetonitrile. petroleum ether 3/10. 510 mg of desired product is obtained (yield 38.5%).

Mass spectrum: m / e = 339 (M + + 1)
1 H-NMR (ppm): 5.04-5.54 (2H, m, vinyl CH); 4.26-4.42 (1H, m, N-CH-CH = CH-); 3.32-3.74 (2H, m, CH2-N ring); 1.12-1.26 (21H, m, tert.C4HgO, 6x CH2); 0.74-0.98 (6H, m, 2 x CH 3).

f / Preparation of 1- (N- (tert.butoxycarbonyl) 2 (S) -aminobutyryl) -2-hexylpyrrolidineamide: Compound XXVII
400 mg (1.18 mmol) of the compound obtained in 100 ml of methanol are dissolved in 100 ml of methanol and 200 mg of 10% palladium on charcoal are added. The mixture is hydrogenated for 2 hours. Chromatography showed that no more starter was left and only one product was formed. The catalyst is removed by filtration and the solvent is removed. 310 mg of an oil are obtained (yield 77.1%).

 Mass spectrum: m / e = 341 (M + +1).

g Preparation of the title compound as oxalate
Cooled to 0 C, 10 ml of trifluoroacetic acid and 300 mg (0.882 mmol) of the compound obtained at f /. The mixture is kept at 00C for one hour. The acid is removed and the residue is dissolved in 100 ml of a solution of
KOH, then extracted with ethyl acetate (2 x 100 ml). The extracts are combined and dried (Na 2 SO 4) and the solvent is then removed. 220 mg of a solid are obtained which is dissolved in ethanol and treated with 82 mg (0.916 mmol) of oxalic acid and ether. By cooling, an inconsistent gel is obtained which leads to 77 mg of a solid by pumping (26.9% reduction).

HPLC (Column: Kromasil C-18 eluent methanol / water / trifluoroacetic acid 40/40 / 0.1): purity = 99.4%
Elemental analysis: C14H28N2O.7 (COOH) 2.O, 7H20.0.2C2HSOH
Found: C, 58.84%; H, 10.13%; N = 8.43%
Calculated: C, 58.81%; H = 9.99%, N = 8.47%
1 H-NMR (400 MHz, DMSO-d 6, ppm): 3.90-4.05 (2H, m,
CH 2 pyrrolidine); 3.55-3.85 (1H, m, NCHCO); 3.3-3.45 (1H, m,
NCH pyrrolidine); 1.35-1.9 (7H, m, 2 x CH 2 pyrrolidine,
CH2butyryl, N-CH-CH (H) -CsH1l); 1.1-1.35 (9H, m, 4 x CH2, N CH-CH (H) -CsH11); 0.8-0.95 (6H, m, 2 x CH3).

Example 13: Preparation of 1- (2 (S) -aminobutyryl) -2 - [(1-oxo) hexyl) pyrrolidinamide (UCL 1420-H); lane 8
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-R7; R7 = n.CsH1l
Preparation of N-benzyl-L-proline
11.5 g (99.8 mmol) of L-proline are suspended in ethanol (200 ml) and 13.8 g (99.8 mmol) of potassium carbonate are treated. potassium iodide (in catalytic amount) and 13.9 g (99.8 mmol) of benzoyl chloride. The mixture is refluxed and thus maintained for 3 hours. It is then stirred at room temperature overnight. The solid formed is filtered off and the filtrate is concentrated. The residue is dissolved in water (200 ml) and the pH is adjusted to 7 with hydrochloric acid.

After removal of the water, the residue is triturated in hot acetone. The solid obtained (yield 65%) is sufficiently pure to stretch used in the next step.

 Mp = 1500C (decomposed).

b / Preparation of 1- [2- (N-benzyl) pyrrolidinyl-1-hexanone: compound XXVIII
Dry (400 ml) nitrogen (20 g, 97 mmol) of N- (benzyl) -L-proline obtained under nitrogen was suspended and cooled in ice. 120 ml of freshly prepared 1.66M pentyllithium are added over 10 minutes and the mixture is warmed to room temperature and stirred overnight. It is then cooled in ice and concentrated sulfuric acid is added. The mixture is basified with sodium bicarbonate and then extracted with methylene chloride. After drying (Na2SO4) extracts and removing the solvent, an oil is obtained.

 1 H-NMR (CDCl 3, ppm): 7.2 (5H, m, Ph); 3.8 (1H, d, PhCH (H)); 3.3 (1H, d, PhCH (H)); 3.0 (2H, m, NCH 2 ring) 2.1-2.5 (4H, m, NCH 2 CH 2 CH 2 cycle); 2.0 (1H, m, NCHCO); 1.01.9 (8H, m, 4 x CH 2 chain); 0.9 (3H, m, CH 3).

 Mass spectrum: m / e = 260 (M + + 1).

c / Preparation of 1- (2-pyrrolidinyl) -1-hexanone
compound XXIX
Dissolved in methanol (50 ml), 0.342 g (1.32 mmol)
of the compound obtained in b / and palladium on charcoal is added
(10%). This mixture is hydrogenated at 40 psi for 5 hours.

The catalyst is removed by filtration and the
solvent, which leads to the desired product.

1 H-NMR (CDCl 3, ppm) 3.2-3.5 (2H, m, NCH 2); 1.0-2.1
(12H, m, 6 x CH 2); 0.8 (3H, m, CH 3).

 Mass spectrum: m / e = 170 (M + + 1).

d / Preparation of 1- (N-benzyloxycarbonyl-2 (S) -amino-butyryl) -2-L (1-oxo) hexyl) pyrrolidinamide: compound XXX
Dissolve in dry tetrahydrofuran (20 ml), under
nitrogen, 0.313 g (1.3 mmol) of N-benzyloxycarbonyl acid
2 (S) -aminobutyric acid prepared as in Example 3a and
cools in ice. 0.15 g (1.3 mmol) of
N-ethylmorpholine and 0.18 g (1.3 mmol) of chloroformate
isobutyl. After stirring for 30 minutes, the compound
obtained at the same time and the reaction mixture is stirred
a night. It is then diluted with methylene chloride,
washed with a citric acid solution (5%, 3 x 100 ml), then dried and the solvent removed, yielding an oil (80% yield).

1 H-NMR (CDCl 3, ppm): 7.25 (5H, m, Ph); 5.20 (1H, m,
NCHCO); 5.10 (2H, s, PhCH 2) 4.30 (1H, m, NCHCO); 4.0 (2H, m, NCH 2 cycle); 0.9-2.1 (20H, m, 7 x CH2, 2 x CH3).

 Mass spectrum: m / e = 389 (M + + 1).

e / Preparation of the title compound as oxalate
0.3 g (0.77 mmol) of the compound obtained in d) are dissolved in methanol and 0.03 g of palladium-on-charcoal (10%, moisture 50%) is added. The mixture is hydrogenated at 40 psi for 4 hours and then the catalyst is filtered off and the solvent is removed. An oil is obtained which is treated with oxalic acid in ethanol and then with ether. The desired product is obtained as a white solid (80% yield) mp = 129-130 ° C.
Elemental analysis: C14H26N2O2. (COOH) 2.2,75H2O
Found: C, 48.61%; H, 8.20%; N = 6.82%
Calculated: C = 48.78%; H, 8.57%; N = 7.11%
1 H-NMR (CD 3 OD, ppm): 4.0 (2H, m, NCH 2); 3.75 (1H, m,
NCHCO); 3.25 (1H, m, NCHCO); 1.95 (6H, m, COCH 2, NCH 2 CH 2 CH 2); 1.3 (2H, m, CH 2butyryl); 0.9-1.2 (14H, m, CH 3 (CH 2) 3,
CH3butyryle).

 Mass spectrum: m / e = 255 (M + + 1).

Example 14: Preparation of 1- (2-amino-2-methyl-propionyl) -L-proline n-butylamide (UCL 1424-A); way 9
R1 = CH3; R2 = CH3; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-NH-R5; R5 = n.C4Hg
a / Preparation of N- (tert.butoxycarbonyl) -2-amino-2-methylpropionic acid: compound IX
50% aqueous dioxan (100 ml) containing triethylamine (15 ml), 4.2 g (0.041 mol) of 2-amino-2-methylpropionic acid are dissolved. The mixture was cooled in ice and treated with 6.4 g (0.026 moles) of 2- (tert.butoxycarbonyl) -2-phenylacetonitrile. The reaction mixture was stirred at room temperature for 60 hours and then the solvent was removed. The residue was treated with water (60 ml) and ethyl acetate (40 ml) and the medium was made basic. using a 4M sodium hydroxide solution. The phases are separated and the aqueous phase is washed with ethyl acetate (75 ml). It is then acidified with a 5% citric acid solution and then extracted with ethyl acetate (200 ml).

The extract is dried (Na 2 SO 4) and the solvent is removed. 3.9 g of desired product are obtained (yield 74%).

Mp 117-119 ° C
1 H-NMR (CDCl 3, ppm): 1.51 (6H, s, 2 x CH 3); 1.42 (9H, s, tert.C4HgO).

 Mass spectrum (FAB): m / e = 204 (M + + 1).

bl Preparation of N- (tert.butoxycarbonyl) -L-proline n.butylamide
N-tert.-butoxycarbonyl-L-proline (5 g, 0.023 moles) was dissolved in dry tetrahydrofuran (30 ml). It is cooled to -10 ° C. and 3 ml (0.023 mol) of isobutyl chloroformate and 2.9 ml (0.023 mol) of N-ethylmorpholine are added. The mixture is stirred at -10 ° C. for 20 minutes and then 5 ml (0.051 mol) of n-butylamine are added. It is stirred for one hour at 0 ° C and then for 18 hours at room temperature. The solvent is removed and the resulting residue is dissolved in ethyl acetate (200 ml). The solution was washed with water (100 ml), 5% citric acid (40 ml), 10% potassium bicarbonate solution (40 ml) and brine (30 ml). It is dried (Na 2 SO 4) and the solvent is removed, which gives an oil.

After crystallization from hexane, 3.53 g of desired product is obtained (yield 56%).

PF = 65-66 ° C
1 H-NMR (CDCl 3, ppm): 4.23 (1H, brs, NCHCO); 3.40 (1H, brs, NH); 3.22 (4H, brs, 2 x NCH2); 1.65 and 1.85 (4H, m, NCH2CH2CH2proline); 1.42 (9H, s, tert.butoxycarbonyl); 1.33 (4H, m, NHCH 2 CH 2 CH 2 butyl); 0.89 (3H, t,
CH3).

 Mass spectrum (EI): m / e = 270 (M +).

c / Preparation of L-Proline N-Butylamide: Compound V
Dissolved in dichloromethane (10 ml), 1 g (3.7 mmol) of the compound prepared in b /. Cool in ice with stirring and add trifluoroacetic acid (5 ml). The reaction mixture is stirred at room temperature for 3 hours and then the solvent is removed and the residual oil is dissolved in 10% potassium carbonate solution (20 ml). Extracted with ethyl acetate (3 x 25 ml) and dried (Na 2 SO 4) the combined organic phases. After removal of the solvent, 0.62 g of an oil (100% yield) is obtained.

1 H-NMR (CDCl 3, ppm) 7.93 (1H, s, NH); 4.45 (1H, m,
NCHCO); 3.23-3.25 (4H, m, 2 x NCH2); 1.96 and 2.36 (4H, m,
NCH2CH2CH2proline); 1.33 and 1.47 (4H, m, NHCH2CH2CH2butyl) 0.89 (3H, t, CH3).

 Mass spectrum (EI): m / e = 170 (M +).

d / Preparation of 1- (N- (tert.butoxycarbonyl) -2-amino-2-methylpropionyl) -L-proline n.butylamide: compound IV
Dissolved in dichloromethane (15 ml) was 0.6 g (3.5 mmol) of L-proline n.butylamide prepared in c /. It is cooled to 0.degree. C. and treated with 0.7 g (3.5 mmol) of the acid obtained in (a) and 0.7 g (3.5 mmol) of dicyclohexylcarbodiimide. The reaction mixture is stirred at room temperature overnight then filtered and the solvent removed. The resulting residue is dissolved in dichloromethane (50 ml) and washed with a solution of 5% citric acid (30 ml), water (30 ml) and 10% potassium bicarbonate solution ( 30 ml). A charcoal discoloration is then carried out and then dried (Na 2 SO 4) and the solvent is removed. 0.28 g (22% yield) of a white solid are obtained.

Mp 198-203 ° C
1 H-NMR (CDCl 3, ppm): 4.64 (1H, m, NCHCO); 3.61 (2H, m,
NCH2butyl); 3.20 (2H, m, NCH2proline); 2.05 (2H, m, CH 2 proline); 1.86 (2H, m, CH 2 proline); 1.64 (3H, s,
Isobutyl); 1.52 (3H, s, 3-isobutyl); 1.42 (9H, s, tert.CHO); 1.37 (4H, NCH 2 CH 2 CH 2 butyl); 0.86 (3H, t, butyl CH3).

 Mass spectrum (EI): m / e = 356 (M + + 1).

e / Preparation of the title compound as hydrochloride
Hydrochloric acid in glacial acetic acid (M, 5 ml) was added to 24 mg (0.06 mmol) of the compound prepared in d /. The mixture is stirred for 2 hours and then evaporated to dryness. 17 mg (yield 89%) of desired product are obtained.

MP = 155-156 ° C
1 H-NMR (DMSO-d 6, ppm): 4.31 (1H, q, NCHCO); 3.63 (2H, t,
NCH2 proline); 3.00 (2H, m, butyl NCH 2); 1.78 and 1.98 (4H, m, NCH 2 CH 2 CH 2 proline); 1.51 (6H, s, C (CH3) 2); 1.25 and 1.35 (4H, m, NCH 2 CH 2 CH 2 butyl); 0.85 (3H, t, CH 3).

 Mass spectrum (EI): m / e = 255 (M +).

Elemental analysis: C13H25N302.0,8HC1.0,9H20.0.2 (C2Hg)
Found: C, 52.40%; H, 9.42%; N = 13.41%
Calculated: C = 52.53%; H, 9.45%; N = 13.32%
Example 15: Preparation of 1- (2 (S) -aminobutyryl) - (4 (S) -fluoro) -L-proline-n-butylamide (UCL 1395-F); lane 3
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = -CH2-CH (cis.F) -; R4 = CO-NH-R5 R5 = n.C4Hg
a / Preparation of the methyl ester of
N-benzyloxycarbonyl-4 (S) -fluoro-L-proline
4.38 g (17.29 mmol) of N-benzyloxycarbonyl-4 (R) -hydroxy-L-proline methyl ester (commercially available from ALDRICH) are dissolved under nitrogen in dichloromethane (90 ml),
SIGMA) and cooled to -70 ° C and then treated dropwise with 3 ml (22.71 mmol) of diethylaminosulfide trifluoride (DAST). The resulting mixture is heated in one hour and stirred for 18 hours. The reaction mixture is then poured into a water-ice mixture containing NaHCO 3, it is extracted with dichloromethane, the extract is dried (MgSO 4) and concentrated under reduced pressure. The yellow oil obtained is purified by chromatography on silica gel. eluting with a mixture of petroleum ether / ethyl acetate 40/60 then 50/50. After evaporation under vacuum, 3.44 g of an oil (yield 70.5%) are obtained; Rf = 0.5 (50/50 petroleum ether / ethyl acetate)
1 H-NMR (CDCl 3, ppm): 7.40-7.27 (5H, m, ArH); 5.22 (1H, dt, -CHF-, HF = 52.4Hz, J = 3.7Hz); 5.14-5.06 (2H, m,
CH 2 benzyl); 4.58 (dd, H, J = 30.1 Hz, J = 9.5 Hz); 3.963.62 (2H, m, -CH2N-); 3.64, 3.75 (3H, OCH3); 2.59-2.47 (1H, m, -CH 2); 2.47-2.24 (1H, m, -CH 2 -).

 Mass spectrum (EI): m / e {% intensity) = 281 (M +, 1.36); 261 (M + HF, O 01); 91 (Ph-CH 2 +, 100).

b / Preparation of N-benzyloxycarbonyl-4 (S) -fluoro
L-proline
3.29 g (11.70 mmol) of the compound prepared in (a) are dissolved in methanol (30 ml) and 7 ml (14 mmol) of 2M sodium hydroxide are added dropwise at -50C. The mixture is then left at room temperature for 17 hours. After addition of water (100 ml), the methanol is removed under vacuum and the aqueous solution is washed twice with dichloromethane. This solution is then acidified to pH = 3 with citric acid (20%) and then extracted with dichloromethane (3 × 50 cm 3). The extracts are dried (MgSO4), filtered and the solvent removed in vacuo (by adding ether at the end). 2.91 g of a white solid are thus obtained (yield 93%).

PF = 123-124C
1 H-NMR (400 MHz, CDCl 3, ppm) 7.38-7.26 (5H, m, ArH) 5.32-5.15 (3H, m, -CHF-RF = 52.16 Hz, CH 2 benzyl) ; 4.61 (1H, dd, alphaHJ = 27.6 Hz, J = 9.6 Hz); 3.98-3.60 (2H, m, CH 2 N); 2.78-2.54 (1H, m, -CH 2 -); 2.52-2.24 (1H, m, -CH 2 -).

TLC (ethyl acetate): Rf = 0.04
Mass Spectrum (EI): m / e (% intensity) = 267 (M +, 1.82); 247 (M + H, 0.02); 91 (PhCH2 +, 100).

c / Preparation of N-benzyloxycarbonyl-4 (S) fluoro-L-proline n-butylamide
1.40 g (5.24 mmol) of the compound obtained in b) are dissolved under argon in dry tetrahydrofuran (40 ml) and 0.66 ml (5.24 mmol) of 4-ethylmorpholine are added at 0.degree. C then 0.685 ml
(5.24 mmol) of isobutyl chloroformate at -5 ° C. A white precipitate of 4-ethylmorpholine hydrochloride is formed.

After stirring for 20 minutes at -5 ° C., 0.54 ml is added.
(5.46 mmol) of n-butylamine and the reaction mixture is stirred for 40 minutes at 0 ° C. and then for one hour at room temperature. The solid is removed by filtration, washed with dry tetrahydrofuran and the resulting solution is evaporated under vacuum. The colorless residue is then treated with 50 ml of a 5% citric acid solution and 120 ml of ethyl acetate. After shaking thoroughly, the phases are separated, the organic phase is washed with 50 ml of 10% KHCO 3 and then with 50 ml of brine. After drying (MgSO 4) and removal under vacuum of the solvent, 1.89 g of white solid which is washed with petroleum ether and dried under vacuum on P2O5. 1.50 g of desired product are obtained in the form of a white solid.

Mp = 79-80 ° C
TLC (ethyl acetate / 50/50 petroleum ether): 0.5
1 H-NMR (400 MHz, CDCl 3, ppm): 7.34 (5H, brs, ArH) 6.50-6.00 (1H, broad, -NH-); 5.21 (1H, dt, -CHF-HF = 51.9
Hz, J = 3.4 Hz); 5.20-5.00 (2H, m, CH 2 benzyl); 4.48 (1H, d, aH J = 1.4 Hz); 4.02-3.38 (2H, m, -CH2N proline); 3.32 to 3.02
(2H, m, -CH2-NH-NHbutyl); 2.90-2.52 (1H, m, -CH2-proline) 2.48-2.08 (1H, m, -CH2-proline) 1.54-1.16 (4H, m, -CH2)
NHbutyl); 0.87 (3H, brs, butyl CH 3 NH).

d / Preparation of 4 (S) -fluoro-L-proline n.butyiamide
1.48 g (4.59 mmol) of the compound obtained in (c) were dissolved in 40 ml of methanol and 0.56 g of palladium on wet activated charcoal (t0%) was added. The reaction mixture was hydrogenated at 60 psi for 3 hours and then the solution was filtered and concentrated in vacuo. An oil is obtained which crystallizes after drying in vacuo over P2O5 for 18 hours, yielding 0.76 g of desired product (88.5% yield).

PF = 30 C
1 H-NMR (400 MHz, CDCl 3, ppm): 7.53 (brs, 1H, -NH-) 5.16 (1H, dt, -CHF-J1 = 53.1Hz, J2 = 3.7HZ) ; 3.82 (1H, dd, alphaH, J1 = 10.3 Hz, J2 = 3.3 Hz); 3.40-3.10 (4H, m, -CH 2 -N- and -CO-N-CH 2 -); 2.48 - 2.20 (2H, m, -CH 2 -proline); 2.12 (1H, brs, -NH-); 1.42-1.52 (2H, m, -CH2-NHbutyl); 1.37-1.28 (2H, m, -CH2-NHbutyl); 0.91 (3H, t, J = 7.34 Hz, -CH 3).

e / Preparation of 1- (N- (tert.butoxycarbonyl) -2 (S) -aminobutyryl) - (4 (S) -fluoro) -L-proline n-butylamide
The procedure is as in step c) starting from 0.81 g (3.98 mmol) of N- (tert.butoxycarbonyl) -2 (S) -aminobutyric acid prepared as in Example 2 a / and using the compound obtained in d / instead of n-butylamine. The resulting crude product has two TLC tasks with similar Rf. These two products are separated by chromatography on silica gel, eluting with a gradient of the mixture of ethyl acetate / petroleum ether from 20/80 to 60/40. 0.74 g of pure amide (second product) which crystallized after drying on P 2 O 5 under vacuum for 5 days (yield 51%).

PF = 67-68 ° C
(trans isomer: T and cis isomer: C)
1 H-NMR (400 MHz, CDCl 3, ppm): 7.52 (0.37 H, brs, -NH)
C); 6.55 (0.63H, brs, -NH-T); 5.31 (0.37H, dt, -CHF-T
J1 = 52.3 Hz, J2 = 4.0 Hz); 5.23 (0.63H, dt, -CHF-C J1 = 50.1
Hz, J2 = 3.2 Hz); 5.19 (0.63H, broad d = -NH-TJ = 8.3Hz) 5.19 (0.37H, broad, -NH-C = 6.7Hz); 4.76 (0.63H, d, aH T
J = 9.5 Hz); 4.43 (0.37H, d, xH C, J = 9.2 Hz); 4.33 (0.63H, q, "HT J1 = 8.5 Hz, J2 = 6.7 Hz); 4.09-3.76 (2.37H, m, -CH2 N-proline and HC) 3.46-2.85 (3H, m, -CO-N-CH 2 and -CHaminobutyryl); 2.34-2.07 (1H, m, -CH 2 aminobutyryl) 1.89-1.20 (15H, m.p. m, tert.C4Hg and -CH2-proline and NHbutyl) 1.00 (1.89H, t, CH3aminobutyryl TJ = 7.3 Hz), 1.00 (1, 11H, t, CH3aminobutyryl CJ = 7.5Hz) 0.89 (3H, t, CH3NHbutyl,
J = 7.3 Hz).

Mass Spectrum (FAB): Found: MH + = 374 (11.7); 58 (100.0); calculated: M = 373
f / Preparation of the title compound in the form of trifluoroacetate
A 50/50 solution of CF 3 COOH / CH 2 Cl 2 is cooled to 0 ° C. and 0.20 g (0.547 mmol) of the compound prepared in the reaction mixture is added. Stir at room temperature for one hour and 15 minutes and then remove the acid and solvent in vacuo to give a yellow oil. 2 x 6 ml of CH 2 Cl 2 are then added and evaporated under vacuum to remove excess acid. The resulting crude product is washed with anhydrous diethyl ether (3 x 20 ml) and a white solid is obtained which is dried for 48 hours at room temperature under 0.25 mmHg. Finally, 0.20 g of desired product is obtained (yield 90%).

PF = 60-63 ° C
(Trans isomer and C: cis isomer)
1 H-NMR (400 MHz, CD 3 OD, ppm): 5.34 (0.73H, dm, -CHF-T) 5.27 (0.27H, dm, -CHF-C); 4.65 (1H, dd, 4-Haminobutyryl) 4.13 (0.73H, dd, α-Hproline T); 4.05-3.63 (2H, m, CH 2 -Nproline and 0.27H, -Hproline C); 3.28-3.08 (2H, m, -N-CH 2 -NHbutyl); 2.68-2.25 (2H, m, -CH 2 -aminobutyryl); 2.10-1.75 (2H, m, -CH2proline); 1.54-1.42 (2H, m, -CH 2 -CH 2 -CH 3); 1.40-1.29 (2H, m, -CH 2 -CH 2 -CH 3); 1.11 (2.19H, t, CH3aminobutyryl T); 1.01 (0.81H, t, CH3aminobutyryl C); 0.94 (0.81H, t, CH3NHbutyl
C); 0.92 (2.19H, t, CH3NHbutyl T).

 HPLC (detector: 215 nm, column: Kromasil C18 250 mm x 4.6 mm, 5 μm, flow rate: 1.0 ml / min, eluent: A = H20 + 0.1% TFA and B = CH30H + 0.1 % TFA, A / B = 70/30, mobile phase): majority peak = 99.9%; retention time = 10.29 minutes.

Mass Spectrum (FAB): MH + found = 274 (100%)
calculated = 273
IR (KBr pellet, cm-1): 3438 (m, amide); 3073 (m,
NH3 +); 1668 (s, C = O).

Microanalysis: C13H2402N3F.1,1CF3COOH
Found: C = 45.90%; H, 6.30%; N = 10.48 9s
Calculated: C = 45.78%; H, 6.34%; N = 10.54%
Example 16: Preparation of 1- (2 (S) -aminobutyryl) - (4 (S) -benzyl) -L-proline-n-butylamide (UCL 1433-H); lane 4
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = -CH2-CH (CH2Ph) -; R4 = CO-NH-R5 R5 = n.CqHg
a / Preparation of the methyl ester of N-benzyloxycarbonyl-4-keto-L-proline
Acetone (reactive grade) (5.0 g, 17.90 mmol) of N-benzyloxycarbonyl-4-hydroxy-L-proline methyl ester (commercially available from ALDRICH, SIGMA) was dissolved in 300 ml of acetone (reagent grade). magnetically, and 15 ml of chromic acid in approximately 8N sulfuric acid are added over a period of about 3 minutes. Stirring is continued for one hour and then the excess oxidant is destroyed by adding 5 ml of isopropanol in 20 minutes. The solvent is removed and the residue is dissolved in ether and filtered through fluorisil to remove the chromium salts. The crude product obtained after evaporation is purified by flash chromatography on silica eluting with a gradient of ethyl acetate / petroleum ether mixture (bp <40 ° C) 15/85 to 40/60. 3.97 g of colorless oil are obtained after drying under reduced pressure overnight (yield 80%).

Rf = 0.85 (ethyl acetate)
1 H-NMR (200 MHz, CDCl 3, ppm) 7.30 (5H, brs, ArH) 5.20-5.00 (2H, m, CH 2 benzyl); 4.85 (1H, t, -H); 3.95 (2H, brs, -CH2-Nproline); 3.70-3.60 (3H, d, -OCH3); 3.05-2.80 (1H, m, -CH 2proline); 3.65-3.50 (1H, m, CH 2 -proline).

Mass Spectrum (FAB): m / e (% intensity) = 300 ((M +
Na) +, 100.0); 278 (MH +, 13.0); 154 (43.5); 136 (12); 107 (12.5); 91 (100).

b / Preparation of the methyl ester of N-benzyloxycarbonyl-4-benzylidene-L-proline
3.68 g (9.46 mmol) of benzyltriphenylphosphonium chloride are suspended in 30 ml of dry tetrahydrofuran and this mixture is added to a solution of 0.22 g (9.65 mmol) of sodium hyride ( 95%) in 20 ml of dry tetrahydrofuran under nitrogen. 25 ml of dry DMSO are then added and the mixture is heated to 70c until it becomes homogeneous (4 hours). After cooling the solution to 50 ° C., a mixture of 2.00 g (7.90 mmol) of the ester obtained in (a) in 10 ml of dry tetrahydrofuran is added over 5 minutes. The reaction mixture is then heated to 70.degree. C. for 16 hours and then poured into 200 ml of water / ice containing 1.4 g of KHCO 3. 150 ml of dichloromethane are then added and the organic phase is then separated off and the aqueous phase is extracted once with 100 ml of dichloromethane. After drying (MgSO 4) of the organic phases and evaporation in vacuo, the crude product obtained is purified by flash chromatography on silica eluting with a mixture of ethyl acetate and petroleum ether 15/85 then 35/65.

After drying in vacuo over P2O5 overnight, 2.19 g of a colorless oil (79% yield) was obtained.

Rf = 0.90 (ethyl acetate)
1 H-NMR (400 MHz, CDCl 3, ppm): 7.50-7.05 (10H, m, ArH) 6.53-6.13 (1H, m, vinyl H); 5.30-5.00 (2H, m, H benzyl) 4.73-4.30 (3H, m, cs-H and N-CH2proline); 3.80-3.50 (3H, dd,
OCH3) 3.33-3.10 (1H, m, -CH2proline); 3.05-2.67 (1H, m, -CH 2proline)
HPLC (column: Kromasil silica gel 250 x 4.6 mm, flow rate 1 ml / min, detector 254 nm, eluent dichloromethane) two isomers (cis and trans) in proportion 44/56; retention time: 9.28 minutes (44%) and 10.42 minutes (56%) total purity of both isomers: 98.3%.

 Mass spectrum (FAB, (MNOBA + NaI matrix): m / e (% intensity) = 300 (MH + + Na, 97), 278 (MH +, 13), 154 (44), 91 (C7H7 +, 100).

c / Preparation of the methyl ester of 4 (S) -benzyl
L-proline
0.95 g of activated palladium on activated carbon (10% wet) is added to a solution of 2.10 g (5.97 mmol) of the ester obtained in b / in 250 ml of methanol and this reaction mixture is hydrogenated. for 2 hours and 30 minutes at 60 psi. After filtration of the solution, concentration in vacuo and drying over P2O5 overnight, 0.889 g of desired product (yield 67%) is obtained.

 The NMR spectrum shows the presence of two diastereoisomers in a 90/10 ci / trans ratio.

(T = trans isomer and C = cis isomer)
1 H-NMR (400 MHz, ppm): 7.32-7.12 (5H, m, ArH); 3.82 (1H, t, aH, J = 8.0 Hz); 3.74 (3H, s, OCH 3) 3.08-3.02 (1H, m, -CH 2 -Nproline); 2.80-2.73 (1H, m, -CH2N proline); 2.72-2.58 (2H, m, Hb-benzyl); 1.60-1.70 (0.1H, m, PhCH 2 CH T); 1.50-1.59 (0.9H, m, PhCH2CH C).

d / Preparation of 1- (N-benzyloxycarhonyl-2 (S) -aminobutyryl) - (4 (S) -benzyl) -L-proline methyl ester
0.66 g (4.01 mmol) of 3-hydroxy-1,2,3-benzotriazine-4 (3H) -one and 0.83 g (4.01 mmol) of dicyiohexylcarbodiimide (DCC) are added at 0 ° C. to 0.degree. solution of 0.815 g (3.43 mmol) of N-benzyloxycarbonyl-2 (S) -aminobutyric acid prepared as in Example 3 a / in a mixture of 5 ml of dimethylformamide and 5 ml of methylene chloride. After one hour and 15 minutes, a mixture of 0.88 g (4.01 mmol) of the obtained ester with 5 ml of CH 2 Cl 2 was added. Stirred at 0 C for 15 minutes and then two days at room temperature. After removing the solvent in vacuo, the residue is taken up in ethyl acetate. Dicyclohexylurea is removed by filtration, the filtrate is washed successively with 4% NaRCO 3, 10% citric acid and then evaporated under vacuum. .

The crude product thus obtained is purified by flash chromatography on silica eluting with a gradient of ethyl acetate / petroleum ether mixture 15/85 to 35/65. After evaporation, 1.30 g of a yellow paste (yield 74%) are obtained.

 1 H-NMR (200 MHz, CDCl 3, ppm): 7.50-6.95 (10H, m, ArH) 5.60 (1H, d, -NH-); 5.05 (2H, m, PhCH2O); 4.50-4.35 (2H, m, a-H); 3.95-3.60 (4H, m, OCH3 and -CH2-Nproline) 3.35-3.20 (1H, m, -CH2-Nproline); 2.80-2.10 (4H, m, PhCH2C and -CH2proline); 1.95-1.50 (3H, m, -CH 2 -aminobutyryl and H-proline); 0.95 (3H, t, CH3aminobutyryl).

Rf = 0.61 (ethyl acetate)
Preparation of 1- (N-benzyloxycarbonyl-2 (S) -amino-butyryl) - (4 (S) -benzyl) -L-proline
1.30 g (2.96 mmol) of the ester obtained in d) are saponified with 1.75 ml (3.50 mmol) of 2M NaOH in 30 ml of methanol for 20 hours at room temperature. After evaporation under vacuum, taken up in 100 ml of distilled water, washed twice with ether and then acidified to pH = 1 with 5N aqueous HCl, extracted three times with 150 ml of ethyl ether, dried (MgSO4). organic phases, filtered and evaporated in vacuo. 1.05 g of desired product is obtained as a white solid (83.5% yield).

Mp = 56-57 ° C
1 H-NMR (200 MHz, CDCl 3, ppm): 7.5-7.0 (10H, m, ArH); 5.7 (1H, d, -NH-); 5.05 (2H, brs, PhCH2O); 4.50-4.25 (2H, m, aH); 4.00-3.80 (1H, m, -N-CH2proline); 3.30-3.10 (1H, m, -N-CH2proline); 2.80-2.10 (4H, m, PhCH2C and -CH2proline) 1.90-1.40 (3H, m, -CH2aminobutyryl and Hproline); 0.90 (3H, t,
CH3aminobutyryle).

f / Preparation of 1- (N-benzyloxycarbonyl-2 (S) -aminobutyryl) - (4 (S) -benzyl) -L-proline n-butylamide
The procedure is as indicated in Example 15c / from the compound obtained previously in c /. The resulting crude product is purified by flash chromatography on silica eluting with a gradient of 15/85 to 50/50 ethyl acetate / petroleum ether. Two samples of the desired product are obtained.

g / Preparation of the title compound in the form of oxalate hydrate
A solution of 0.75 g of the compound obtained above in methanol was hydrogenated at 60 psi for 2 hours and 30 minutes using 0.30 g of 10% palladium on charcoal (wet). After removal of the catalyst by filtration and of the solvent in vacuo, the crude product obtained is purified by flash chromatography using, as first eluent, a mixture of ethyl acetate / petroleum ether 50/50 and then a mixture of ethyl acetate / methanol 50 / 50. After evaporation of the solvent, the resulting oil is taken up in isopropanol and filtered to remove the silica. 350 mg (1.01 mmol) of free base are obtained. 0.12 g (1.32 mmol) of oxalic acid in ethanol is added and ether is added. After crystallization, 0.18 g of desired product is obtained (yield 25%). HPLC analysis indicates two isomers which are considered cis- and trans- (4-benzyl) -1-proline diastereoisomers in a 90/10 cis / trans ratio.

PF = 89-93 ° C (open capillary)
1 H-NMR (400 MHz, CD3OD, ppm): 7.32-7.15 (5H, m, ArH) 4.48-4.28 (1H, m, aH); 4.22-4.10 (1H, m, aH); 3.87-3.68 (1H, m, -CH2-Nprolin); 3.42-3.06 (3H, m, -CH2Nproline and -CH2-N NH2butyl); 2.84-2.44 (3H, m, -CH2Ph and CH2proline); 2.36-2.10 (1H, m, -CH2proline); 2.02-1.76 (2H, m, -CH2aminobutyryl); 1.67-1.26 (5H, m, -CH2CH2NH2butyl and Hproline); 1,12-0,86 (6H, m, -CH3aminobutyryl and -CH3
NH2butyle).

HPLC (column: Kromasil C18 250 mm x 4.6 mm, 5 μm, UV detector: 215 nm, flow rate: 1 ml / min; mobile phase A =
CH3CN and B = H20 + 0.1% TEA, A / B = 30/70): majority peak = 88.92%, retention time 12.63 minutes (cis isomer) 10.08%, retention time 14, 93 minutes (trans isomer); minority peak = 0.99%, retention time 3.3 minutes.

Microanalysis: C20H31N2O2.1, O5 (COOH) 2.O, 9H20
Found: C, 58.19%; HH = 7.24%; N = 8.95 8
Calculated: C, 58.18%; H, 7.71%; N = 9.21%
Mass Spectrum (FAB): m / e (% intensity) = 368 (M + + Na, 14); 346 (M +, 100); 261 (68); 160 "(47); 91 (C7H7 +, 7).

Example 17 Preparation of 1- (2 (S) -aminobutyryl)
L-thiaprolineamide (UCL 1374-F); lane 1
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = -CH2S-; R4 = CO-NH-R5; R5 = H
The procedure is as in the preparation examples according to route 1 given previously. The desired compound is obtained in the form of trifluoroacetate.

PF = 97-105 ° C
1 H-NMR (400 MHz, DMSO-d6, ppm): 4.50-5.0 (2H, dd, NCH2S); 4.71 (1H, t,? - chaminobutyryl); 4.36 (1H, m, NCHCO cycle); 3.0 (2H, dd, S-CH 2 -CH); 1.77 (2H, m, CH 3 CH 2 -); 0.92 (3H, m, CH 3 -).

IR (KBr, cm-1): 3399 (m, -NH 2); 2977 (m, -CH); 1672 (s,
C = O); 1640 (s, C = O).

Microanalysis: C8H15N3O2S.1,0CF3COOH.0,2H2O
Found: C, 35.84%; H, 5.07%; N = 12.04%
Calculated: C, 35.63%; H, 5.07%; N = 11.98%
Example 18: Preparation of 2- (2 (S) -aminobutyryl) -1,2,3,4-tetrahydro-3 (S) -isoquinolinecarboxylic acid n-butylamide (UCL 1421-H); lane 3
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 =

R4 = CO-NH-R5; R5 = n.c4Hg
Preparation of 1,2,3,4-tetrahydro-3 (S) -isoquinolecarboxylic acid
5.0 g (30.27 mmol) of L-phenylalanine are dissolved in 50 ml of concentrated hydrochloric acid and 4.0 g of parafomraldehyde and 1 ml of concentrated sulfuric acid are added. two days. After cooling and filtration, the resulting solid is dissolved in a hot water / ethanol mixture: 1/1 (v / v) and added
Aqueous NH4OH (30%) until pH 7.0 is reached. The crystals which precipitated after cooling the solution are recovered by filtration, washed with ethanol and dried. 2.68 g of desired product are thus obtained (yield 50%).

PF = 315C (lit.) 23
Mass Spectrum (FAB): m / e (% intensity) = 178 (MH +, 23); 154 (100).

b / Preparation of N-benzyloxycarbonyl-1,2,3,4-tetrahydro-3 (S) -isoquinolinecarboxylic acid
An ice-water bath, a solution of 2.50 g (14.11 mmol) of the acid prepared in 2N NaOH (8 ml) was cooled and stirred vigorously with a magnetic stirrer. 2.21 ml (15.52 mmol) of benzyl chloroformate and 8 ml of 2N NaOH are added alternately in approximately 10 portions. The temperature of the reaction mixture is maintained between 0 and 5 ° C. by controlling the rate of addition of the reagents. 9 ml of 2N NaOH are added and the mixture is kept at room temperature for 30 minutes. The alkaline solution is washed with ether (4 × 50 ml) and then acidified by addition of 5N HCl (pH <2). The mixture is then extracted with methylene chloride, dried over MgSO 4, filtered and evaporated in vacuo. empty. After drying the resulting solid on P 2 O 5, 3.73 g of desired product are obtained (yield 85%).

Mp = 52-55 ° C
1 H-NMR (200 MHz, CDCl 3, ppm): 7.55-7.00 (9H, m, Ar-H) 5.10 (2H, d, benyl H); 5.00-4.50 (3H, m, "" -H and -CH2-
Ntetro-hydroisoquinoline); 3.20-3.00 (2H, m, -CH2tetrahydroisoquinoline).

 Mass Spectrum (FAB): m / e (% intensity) = 334 ((M + Na) +, 0.1); 312 (MH +, 3.5); 91 (C7H7 +, 100).

Preparation of N-benzyloxycarbonyl-1,2,3,4-tetrahydro-3 (S) -isoquinolinecarboxylic acid n-butylamide
The procedure is as in Example 15 c / from the acid obtained in b /. The reaction mixture is stirred at 0 ° C. for 40 minutes and then at room temperature overnight. 100 ml of methylene chloride are then added and the solution is washed with 3 x 50 ml of 10% citric acid. The phases are separated after shaking vigorously and then the organic phase is washed with 2 x 50 ml of 5% NaHCO3. After drying on
MgSO 4, the crude product obtained is purified by flash chromatography eluting with a mixture of ethyl acetate / petroleum ether 70/30 (v / v). After evaporation under vacuum, the resulting product is dried over P 2 O 5 overnight. 2.36 g of pure amide are thus obtained in the form of an oil (yield 57%).

 1 H-NMR (200 MHz, CDCl 3, ppm) 7.45-7.05 (9H, m, Ar-H) 5.80 (1H, d, -NH-); 5.25-5.06 (2H, m, -CH 2-Ph); 4.90-4.40 (3H, m, -H and -NH-CH2tetrahydroisoquinoline); 3.42-2.78 (4H, m, -NH-CH2NH2butyl and -CH2tetrahydroisoquinoline); 3.60-3.40 (2H, m, -CH2tetrahydroisoquinoline); 1.18-0.66 (7H, m, -CH2CH2CH3 NHbutyl).

Rf (ethyl acetate) = 0.60
Mass Spectrum (FAB): m / e (% intensity) = 367 (MH +, 13.9); 91 (C6H5 +, 100).

d / Preparation of 1,2,3,4-tetrahydro-3 (S) isoquinolinecarboxylic acid n-butylamide
Hydrogenation of the compound obtained under c /, under the conditions of Example 15 d /. 1.30 g of desired product are obtained (yield 89%).

Mp = 58-60 ° C
1 H-NMR (400 MHz, CDCl 3, ppm): 7.26-7.02 (5H, m, Ar-H and -NH amide); 4.08-3.98 (2H, m, α-H and -CH 2 Ntetrahydroisoquinoline) 3.51-3.58 (1H, m, -CH 2 -NHetrahydroisoquinoline); 3.32-3.20 (3H, m, -CH 2 amide and 1H-CH 2 tetrahydroisoquinoline); 2.76-2.88 (1H, m, -CH2tetrahydroisoquinoline); 1.81 (1H, brs, -NHtetrahydroisoquinoline); 1.55-1.46 (2H, m, -NH-CH2-CH2-CH2-CH3); 1.401.29 (2H, m, -NH-CH2-CH2-CH2-CH3); 0.93 (3H, t, CH 3 amine
J = 7.3 Hz).

e / Preparation of 2- (N-benzyloxycarbonyl-2 (S) -aminobutyryl) -1,2,3,4-tetrahydro-3 (S) -isoquinoline carboxylic acid n-butylamide
The procedure is as in step c /, from N-benzyloxycarbonyl-2 (S) -aminobutyric acid prepared as in Example 3 a / and from the compound obtained at dl. The reaction mixture was allowed to stand overnight at room temperature, then 40 ml of methylene chloride was added and this solution was washed with 3 x 50 ml of 10% citric acid. After shaking vigorously, the phases are separated and the organic phase is washed with 2 × 50 ml of 5% NaHCO 3. After drying over MgSO 4 and evaporation in vacuo, the resulting crude product is purified by flash chromatography on silica eluting with a mixture. ethyl acetate 15/85 then 35/65. 0.63 g of desired product (yield 49.5%) is obtained in the form of an oil which is dried under vacuum over P2O5 overnight.

Mass Spectrum (FAB): m / e (% intensity) = 474 ((M +
Na) +, 0.1); 452 (M +, 2.5); 91 (C7H7 +, 100).

1 H-NMR (400 MHz, CDCl 3, ppm): 7.42-7.08 (9H, m, Ar-H) 6.03 (1H, brs, -NH); 5.68 and 5.39 (1H, d, -NHamide) 5.20-4.88 (3H, m, Hb-benzyl and CH); 4.86-4.38 (3H, m, -CH2 Ntetrahydroisoquinoline and -H); 3.68-2.90 (4H, m, -CH2NNH2butyl and -CH2tetrahydroisoquinoline); 2.00-1.60 (2H, m, -CHaminobutyryie); 1.44-1.16 (2H, m, -CH 2 -CH 2 -CH 2 -CH 3) 1.16-0.96 (2H, m, -CH 2 -CH 2 -CH 2 -CH 3); , 86 and 0.78 (3H, t,
CH3aminobutyryle).

f / Preparation of the title compound as oxalate
Hydrogenation of the compound obtained in e /, under the conditions of step dl. 0.16 g (1.78 mmol) of oxalic acid are then added to the filtered solution and this solution is concentrated in vacuo. Ethanol and then ether (ratio 1/50) are then added, which leads to a crystallized product. After filtration and washing with diethyl ether, the solid is dried over P 2 O 5 under vacuum at 30 ° C. for 24 hours and 0.35 g of desired compound is obtained (yield 60.5%).

PF = 105-110 C (open capillary)
1 H-NMR (400 MHz, CD 3 OD, ppm): 7.34-7.15 (4H, m, Ar-H) 4.86-4.54 (4H, m, 2H and -CH 2 Ntetrahydroisoquinoline); 3.43- 2.88 (4H, m, -CH2-tetrahydroisoquinoline and -NH-CH2-NHbutyl) 2.14-1.80 (2H, m, -CHaminobutyryl); 1.48-1.18 (4H, m, -CH 2
CH2 -NHbutyl); 1.0-1.2 (3H, t, CH3aminobutyryl); 0.7-0.95 (3H, t, CH3 -NHbutyl).

 Mass Spectrum (FAB): m / e (% intensity) = 318 (MH +, 96.1); 245 (30.9); 233 (40.3); 149 (100.0); 132 (65.4); 71 (38.8); 58 (53.7); 57 (73.0).

 HPLC purity (Kromasil C-18 water / methanollacid trifluoroacetic 50: 50: 0.1): 98.6%.

 IR (KBr pellet, cm-1): 3072 (m, NH); 2956 (s, CH) 1652 (s, C = O).

Microanalysis: C18H27N3O2.1,05 (COOH) 2.0,5H2O
Found: C, 57.63%; H, 7.08%; N = 9.53%
Calculated: C, 57.35%; H = 7.21%; N = 9.98%
EXAMPLE 19 Preparation of 1- (2 (S) -aminobutyryl) 2 (R / S) -perhydroazepinecarboxylic acid butylamide (UCL 1482-H) 11
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 4-; R4 = CO-NH-R5; R5 = n.C4Hg
a / Preparation of N-benzyloxycarbonyl-2- (S) -aminobutyric acid 3,4-dihydro-4-oxo-1,2,3-benzotriazine compound XXXVI
2 g (8.43 mmol) of N-benzyloxycarbonyl-2 (S) -aminobutyric acid, prepared as in Example 3a, are dissolved in tetrahydrofuran at 0.degree. 1.741 g (8.43 mmol) of dicyclohexylcarbodiimide and then 1.378 g (8.43 mmol) of 3-hydroxy-1,2,3-benzotriazine-4 (3H) -one are added. The mixture is stirred at room temperature for 1 hour. 3 hours then the solvent is removed. Crystallization from ethyl acetate / petroleum ether 1/1 gave 2.344 g of desired product (yield 72%) as pale yellow crystals.

 1 H-NMR (200 MHz, CDCl 3, ppm) # 8.3 (1H, dArCH); 8.2 (1H, d, ArCH); 8.0 (1H, dt, ArCH); 7.8 (1H, dt, ArCH); 7.3 (5H, m, ArCH); 5.3 (1H, broad d, NH); 5.1 (1H, s, PhCH 2) 4.8 (1H, q, NCHCO); 2.1 (2H, m, CH 2); 1.1 (3H, t, CH 3).

b / Preparation of 2 (R / S) -azepinecarboxylic acid
6 g (4.4 mmol) of (7S, 10 R) -10- (tert-butyl) -9-methyl-1,9-diazobicyclo [5,3,0] decanate are dissolved in a 6M HCl in a bomb. -8-one prepared according to the method of Seebach et al.

(Annalen der Chemie, 1215-1232 (1989)) and heated at 1800C for 3 hours. The mixture is then cooled and extracted with methylene chloride and then the aqueous phase is concentrated. Preparative HPLC (Kromasil C-18, eluent water / methanol / trifluoroacetic acid 80: 20: 0.1) gives 0.4 g of desired product (yield 34%).

 1 H-NMR (200 MHz, DMSO-d6, ppm): 9.0 (1H, OH); 4.0 (1H, dd, NCHCO); 3.1 (2H, m, CH 2); 2.1 (1H, m, CH (H)); 1.6 (7H, m, CH (H), 3 x CH2).

 Mass spectrum: m / e = 144 (M + + 1).

c / Preparation of 1- (N-benzyloxycarbonyl 2 (S) -aminobutyryl) -2 (R / S) -perhydroazepinecarboxylic acid: compound
XXXVI I
0.630 g (1.64 mmol) of the ester prepared was cooled to 0 ° C. in 3 ml of dimethylformamide. 0.308 g (1.198 mmol) of the acid obtained was added to the bl in dimethylformamide (2 ml) containing 0.2 ml). This mixture is stirred at ambient temperature for 5 days and then the solvent is evaporated and the product obtained is chromatographed on silica gel, eluting with a 5: 4: 1 petroleum ether / ethyl acetate / acetic acid mixture. 0.433 g of desired product is obtained (yield 100%).

1 H-NMR (200 MHz, CDCl 3, ppm): 7.25 (5H, m, Ph); 6.0 (1H, d, NH); 5.0 (2H, q, PhCH2); 4.8 (1H, m, NCHCO); 4.5 (1H, m,
NCHCO); 3.6 (1H, m, NCH (H)); 3.1 (1H, m, NCH (H)); 2.3 (1H, m, CH 3 CH (H)); 1.6 (7H, m, CH 3 CH (H), 3 x CH 2); 1.2 (2H, m,
CH2); 0.8 (3H, m, CH 3).

dl Preparation of 1- (N-benzyloxycarbonyl2 (S) -aminobutyryl) -2 (R / S) -perhydroazepinecarboxylic acid n-butylamide of: compound XXXVII I
Dry acid tetrahydrofuran (0.45 g, 1.19 mmol) was dissolved in dry tetrahydrofuran (10 ml) and cooled to -10 ° C. under nitrogen. 0.246 ml (2.08 mmol) of N-ethylmorpholine and 0.26 ml (2.08 mmol) of isobutyl chloroformate are added and the mixture is stirred for 30 minutes. 0.2 ml (2.08 mmol) of n-butylamine is then added and stirring is continued for 3 hours. The solvent is removed and the resulting product purified on silica gel chromatography column eluting with 7: 3 petroleum ether / diethyl ether then 2: 3. The desired product is obtained in 52% yield.

 1 H-NMR (200 MHz, CDCl 3, ppm): 7.3 (5H, m, Ph); 6.4 (1H, broad d, NH); 5.3 (1H, broad d, NH); 5.0 (2H, t, PhCH2); 4.7 (1H, m, NCHCO); 4.5 (1H, m, NCHCO); 3.1 (4H, m, 2 x NCH 2) 0.8-1.8 (20H, m, 7 x CH 2, 2 x CH 3).

 Mass Spectrum (FAB): m / e 418 (M + + 1).

e / Preparation of the title compound as oxalate
Dissolved in methanol (15 ml) containing 0.1 g of 10% palladium on charcoal, 0.25 g (0.42 mmol) of the acid prepared in d / and hydrogenated at 30 psi for 2.5 hours.

After removal of the catalyst and concentration of the solution, an oil is obtained which is taken up in methanol and treated with 0.15 g of oxalic acid. Upon addition of ether, the desired product is precipitated. 0.1 g is obtained (yield 438).

1 H-NMR (200 MHz, CD 3 OD, ppm): 8.0 (1H, broad d, NH); 4.7 (1H, dd, NCHCO); 4.4 (1H, m, NCHCO); 3.8 (1H, m, NCH (H)) 3.5 (1H, m, NCH (H)); 3.2 (2H, m, NCH 2); 2.2 (2H, m, CH 2)
1.8 (6H, m, 3 x CH 2); 1.4 (6H, m, 3 x CH 2); 0.9 (6H, dt, 2 x
CH3).

 Mass Spectrum (FAB): m / e = 289 (M + + 1).

Purity (Kromasil C-18, eluent: methanol / water / trifluoroacetic acid 40: 60: 0.1): 99.3%
(diastereomeric ratio: 3: 2).

Example 20: Preparation of 1- (2 (S) -aminobutyryl) 2 (R / S) -pipecolineamide (UCL 1398-F); lane 4
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 3-; R4 = CO-NH-R5; Rs = H
The procedure is as for the other compounds prepared according to route 4.

PF = 46-50 ° C
1 H-NMR (400 MHz, CD3 OD, ppm): 5.19 (1H, d, Hipipecoline); 4.58 (1H, q, Haminobutyryl); 3.82-3.72 (1H, m, N-CH 2 pipecoline); 3.52-3.42 (1H, m, N-CH2pipecoline); 2.82-2.26 (1H, m, -CH 2 pipecoline); 2.24 - 2.25 (1H, m, CH 2 -pipecoline) 2.20-1.25 (8H, m, CH 2 -pipecoline and aminobutyryl, NH 2 amide) 1.04 (3H, t, CH 3).

Microanalysis; C10H19N3O2.1,1CF3COOH.0,5H2O
Found: C, 42.21%; H = 5.9%; N = 11.72%
Calculated: C, 42.14%; H, 6.12%; N = 12.08%
Example 21: Preparation of 1- (2 (S) -aminobutyryl) 2 (R / S) -azetidinecarboxamide (UCL 1373-F); lane 1
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = -CH2-; R4 = -CO-NH-R5; R5 = H
The procedure is as for the other compounds prepared according to route 1.

PV = 60-64C
1 H-NMR (400 MHz, DMSO-d 6, ppm): 7.05 (2H, brs, NH 2) 4.47 (1H, dd, amino-butyryl); 4.07 (1H, m, aCHcycle); 3.73 (2H, t, N-CH 2); 2.00 (2H, m, CH 3 CH 2); 1.59 (2H, m, NCH 2 CH 2) 0.79 (3H, m, CH 3).

 IR spectrum (KBr pellet, cm-1): 3364 (m, NH); 1674 (s, 2 x C = O); 1510-1439 (m, NH3 +).

Microanalysis: C8H15N3O2.1,1CF3COOH.0,5H2O
Found: C, 38.18%; H, 5.62%; N = 13.08%
Calculated: C, 38.33%; H, 5.39%; N = 13.15%
Example 22: Preparation of 1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid n-butylamide (UCL 1397-H); lane 3 R1 = CH2CH3; R 2 = H; n = 0 and m = 1; R3 = (R6 = H)

R4 = CO-NH-R5; R5 = n.C4Hg
a / Preparation of N-benzyloxycarbonyl 2 (S) -indolinecarboxylic acid n-butylamide
Tetranhydrofuran (150 ml) under nitrogen was dissolved in N-benzyloxycarbonyl (S) -indolinecarboxylic acid (7.699 g, 25.9 mmol), prepared as in Example 24 a /. 2.986 g (25.9 mmol) of N-ethylmorpholine and 3.54 g (25.9 mmol) of isobutyl chloroformate are added at 0 ° C. After 45 minutes, 1.903 g (25.9 mmol) of n-ethylmorpholine are added. butylamine and the suspension is stirred overnight. Methylene chloride (300 ml) is added and the organic phase is washed with 10% citric acid (3 x 100 ml) then NaHCO 3 (3 x 100 ml).

After drying and evaporation, a white solid is obtained which is recrystallized from methanol (yield 80%).

PF 164-1650C
1 H-NMR (CDCl 3, ppm): 6.9-7.8 (9H, m, Ar-H); 5.2 (2H, m,
CH 2 benzyloxycarbonyl); 4.9 (1H, m, alphaH); 2.9-3.8 (4H, m,
CH2cycle CH2N); 1.0-1.5 (4H, m, CH 2 CH 2 CH 2 N); 0.8-1.0 (3H, m, CH 3).

b / Preparation of 2 (S) -indolinecarboxylic acid n.butylamide
The compound prepared in methanol is dissolved in methanol and palladium on charcoal (10%, wet, 10% by weight) is added.

The mixture is hydrogenated for 4 hours at 40 psi in a Parr apparatus. After removal of the catalyst and evaporation of methanol, sufficiently pure crystals are obtained without recrystallization (80% yield).

PF = 110-1110C
1 H-NMR (CDCl 3, ppm): 6.8-7.1 (4H, m, Ar-H); 4.3 (1H, m, H); 2.9-3.5 (4H, m, CH 2 CH 2 CN); 1.0-1.9 (4H, m, CH 2 CH 2 CH 2 N); 0.9 (3H, t, CH 3).

c / Preparation of 1- (N-benzyloxyzarbonyl-2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid n-butylamide
The procedure is as in Example 20 using 0.646 g (2.73 mmol) of N-benzyloxycarbonyl-2 (S) -aminobutyric acid prepared as in Example 3a and 0.595 g (2.73 mmol). ) amide prepared in step b /. An oil is obtained which is recrystallized from CCl4 (yield 65%).

Mp 178-179 ° C
1 H-NMR (CDCl 3, ppm): 7.0-7.2 (9H, m, Ar-H); 4.9-5.3 (4H, m, 2 x &alpha; H, CH 2 benzyloxycarbonyl); 3.5 (2H, m, CH 2cycle); 3.1 (2H, m, CH 2 N); 1.0-2.0 (9H, m, CH 2 CH 2 CH 2 N CH 2 aminobutyryl CH 3 aminobutyryl); 0.9 (3H, m, CH3butyl).

d / Preparation of the title compound in the form of oxalate hydrate
The resulting compound is hydrogenated in the same manner as in Example 5 for N-benzyloxycarbonyl-2 (S) -aminobutyryl-L-proline [2 (S) -methyl] n-butylamide and the title compound is isolated. in the form of oxalate salt. It is sufficiently pure without recrystallization (80% yield).

PF = 142 C
1 H-NMR (CD30D, ppm): 8.25 (1H, m, NH); 7.25 (4H, m, Ar
H); 5.1 (2H, m, 2 x H (concealed by the colvent)); 3.8 (2H, m,
CH2Ar); 3.5 (2H, m, CH 2cycle); 3.2 (2H, m, CH 2 N); 2.0 (2H, m, CH 2 aminobutyryl); 0.9-1.8 (10H, m, CH3aminobutyryl)
CH3CH2CH2CH2N).

 HPLC purity (Lichrosorb R. select b methanol / water 40/60 + 0.1% CF3COOH): 99.81%; retention time = 25.00 minutes.

Analysis: C17H25N3O2 .C2H204.0.75 H20
Find: C = 56.25; H = 6.99%; N = 9.84%
Calculated: C = 56.08 z H = 7.06% N = 10.33%
Example 23: Preparation of 1- (2 (S) -aminobutyryl) 2 (S) -indolinecarboxylic acid n propylamide (UCL 1413-H); lane 3
R1 = CH2CH3; R2 = H; n = 0 and m = 1; R3 =

(R6 = H); R4 = CO-NH-R5; Rs = n.C3H7
a / Preparation of N-benzyloxycarbonyl 2 (S) -indolinecarboxylic acid n.propylamide
The procedure is as in Example 22 a / from the acid
N-benzyloxycarbonyl-2 (S) -indolinecarboxylic acid prepared as in Example 24 a / and n-propylamine. A white solid is obtained which is recrystallized in CCl4 (70% yield).

Mp 185-186 ° C
1 H-NMR (CDCl 3, ppm): 7.0-7.25 (9H, m, Ar-CH); 5.20 (2H, s, Ph-CH 2); 4.90 (1H, m, NCHCO); 3.90 (2H, m, CH2indoline) 3.0-3.5 (NCH2propyl); 0.9-1.8 (5H, m, CH 3 CH 2).

b / Preparation of 2 (S) -indolinecarboxylic acid n.propylamide
Hydrogenation of the compound obtained in step a / under the conditions of its n-butylamide analog of Example 22 b /. After evaporation of the methanol, a white solid is obtained which is sufficiently pure without recrystallization (yield 90%).

1 H-NMR (CDCl 3, ppm) 6.85-7.2 (4H, ArCH); 4.3 (1H, m,
NCHCO); 2.9-3.8 (4H, m, NCH2, CH2indoline); 1.5 (2H, m,
NCH2CH2); 0.9 (3H, t, CH 3).

c / Preparation of 1- (N-benzyloxycarbonyl-2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid n.propylamide
0.439 g (1.85 mmol) of N-benzyloxycarbonyl-2 (S) -amino-butyric acid, prepared as in Example 3a, are dissolved under nitrogen in tetrahydrofuran (50 ml) and cooled in an oven. ice / water bath. 0.213 g (1.85 mmol) of N-ethylmorpholine and then 0.253 g (1.85 mmol) of isobutyl chloroformate are added. After 45 minutes, 0.378 g (1.85 mmol) of the compound obtained in step b / are added and the stirring is maintained for 12 hours. The suspension is poured into CH 2 Cl 2 (100 ml), washed with water and dried. 10% citric acid (3 x 100 ml) then with 5% NaRCO3 (3 x 100 ml). After drying and evaporation, a white solid is obtained which crystallizes in CCl 4 (yield 89%).

Mp 176-177 ° C
1 H-NMR (CDCl 3, ppm): 7.0-7.25 (9H, m, H-Ar); 5.0 (2H, s, CH 2 benzyloxycarbonyl); 4.8 (1H, m, zH); 4.1 (1H, alphaH); 3.3 (2H, m, CH 2cycle); 3.0 (2H, m, CH 2 N); 1.0-2.0 (4H, m,
CH3CH2CH2NCH2aminobutyryl); 0.70-1.00 (CH3aminobutyryl CH3propyl).

dl Preparation of the title compound as oxalate hydrate
Hydrogenation of the compound from step c / under the conditions of Example 5 c / for N-benzyloxycarbonyl-2 (S) -aminobutyryl-L-proline [2 (S) -methyl)] n .butylamide. The desired product is obtained in the form of its oxalate salt which is sufficiently pure without recrystallization (90% yield).

PF = 130-131C
1 H-NMR (CD30D, ppm): 7.70 (1H, m, NH); 6.55-6.90 (4H, m,
H-Ar); 4.20-4.60 (2H, m, 2 x zH); 2.90-3.40 (2H, m, CH 2 cycle); 2.50 - 2.80 (CH 2 N); 1.40-1.80 (2H, m, CH 2 CH 3 propyl) 1.00 (2H, m, CH 2 aminobutyryl); 0.40-0.70 (6H, m,
CH3aminobutyryl CH3propyl).

 HPLC purity (Lichrosorb r.p.select b; CH3OH / H2O + 0.1% CF3COOH): 99.03%; retention time = 12.80 minutes.

Microanalysis: C16H23N302. (COOH) 2.1.25H2O
Found: C, 53.79%; H = 6.90 *; N = 10.45%
Calculated: C, 53.84%; H, 6.68%; N = 10.62%
EXAMPLE 24 Preparation of 1- (2 (S) -aminobutyryl) 2 (S) -indolecarboxylic acid methylamide (UCL 1414-H); lane 3 R1 = CH2CH3; R2 = H; n = 0 and m = 1; R3 = (R6 = H); R4 = CO-NH-R5; R5 = CH3

a / Preparation of N-benzyloxycarbonyl2 (S) -indolinecarboxylic acid
A solution of 20 g (122.56 mmol) of 2 (S) -indoline carboxylic acid in 2N NaOH (62 ml) was cooled in an ice-water bath and stirred with a strong magnetic stirrer. 0 ml (140.09 mmol) of benzyl chloroformate and 68 ml of 2N NaOH in about 10 portions over 60 minutes. The temperature of the reaction mixture is maintained between 0 and 5 ° C. by controlling the rate of addition of the reagents. 50 ml of NaOH are then added and the mixture is kept at room temperature for 30 minutes. The alkaline solution was washed with ether (4 x 150 ml) and then acidified by addition of HCl SN (80 ml, pH <2). A crystalline mass precipitates overnight at room temperature. It is filtered and washed with distilled water (500 ml). After drying over P205, 36.00 g of desired product are obtained (yield 99%).

PF = 117-118 ° C
1 H-NMR (400 MHz, CDCl 3, ppm): 8.30-6.90 (10H, m, ArH and
COOH); 5.45-5.14 (2H, m, CH 2 -Ph); 5.08-4.92 (1H, m, uH) 3.65-3.46 (1H, m, CH2indoline); 3.30-3.15 (1H, m,
CH2indoline).

 Mass Spectrum (FAB) m / e (t intensity) = 342 ((M-H + 2Na) +, 0.2); 320 ((M + Na) +, 4.3); 298 (MH +, 5.7); 297 (M +, 9.3); 91 (C7H7 +, 100).

b / Preparation of N-benzyloxycarbonyl2 (S) -indolinecarboxylic acid methylamide
4.00 g (13.45 mmol) of the acid obtained in a / are dissolved in 90 ml of dry tetrahydrofuran and 1.71 ml (13.45 mmol) of 4-ethylmorpholine at 0 ° C. is added. solution at -5 ° C. and 0.62 ml (4.78 mmol) of isobutyl chioroformate are added dropwise. After 30 minutes at -5 ° C., the solution is saturated with methylamine (gas). The mixture is left overnight at room temperature and then 150 ml of dichloromethane are added and this solution is washed with 3 x 150 ml of 10% citric acid. After shaking vigorously, the phases are separated, the organic phase is washed with 2 × 150 ml of 5% NaHCO 3. After drying over MgSO 4, the crude product is purified by washing with hot CCl 4. 2.94 g of a white solid corresponding to the pure amide (yield 70.5%) are obtained.

Mp 210-211 ° C
1 H-NMR (400 MHz, CDCl 3, ppm): 7.75 (1H, brs, COOH) 7.45-6.95 (9H, m, Ar-H); 5.90 (1H, brs, NH); 5.37-5.10 (2H, m, CH2-Ph); 4.93 (1H, dd, ci-H) 3.62-3.16 (2H, m,
CH2indoline); 2.71 (3H, s, CH 3).

 Mass Spectrum (FAB): m / e (% intensity): 311 (MH +, 11.3); 310 (M +, 11.4); 91 (C7H7 +, 100).

c / Preparation of N-benzyloxycarbonyl2 (S) -indolinecarboxylic acid methylamide
A solution of 2.91 g (9.38 mmol) of the compound obtained in step b / in 120 ml of methanol is hydrogenated under 60 psi for 3 hours in the presence of 1.20 g of 10% palladium on activated carbon ( wet). After filtration and concentration in vacuo, the resulting oil crystallizes after drying overnight over P2O5.

1.50 g is obtained (yield 91%).

Mp = 125-130 ° C
1 H-NMR (200 MHz, CDCl 3, ppm): 7.22-7.03 (3H, m, NH 2 O and 2 ArH); 6.82 (1H, t, J = 7.4 Hz, ArH); 6.73 (1H, d, J = 7.8 Hz, ArH); 4.42 (1H, t, -HJ = 9.4 Hz); 4.14 (1H, brs, NH); 3.65 - 3.55 (1H, m, CH 2); 3.14-3.02 (1H, m, CH 2) 2.84 (3H, d, J = 15.0Hz, CH 3).

 Mass Spectrum (FAB): m / e = 177 (MH +, 42.4); 176 (M +, 33.2); 118 (100.0).

d / Preparation of 1- (N-benzyloxycarbonyl-2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid methylamide
1.73 g (8.51 mmol) of N- (tert.butoxycarbonyl) -2 (S) -amino-butyric acid, prepared according to Example 2a, were dissolved under argon in 40 ml of dry tetrahydrofuran and 1.07 ml (8.51 mmol) of 4-ethylmorpholine at 0 ° C. and then 1.11 ml (8.51 mmol) of isobutyl chloroformate are added at -5 ° C. A white precipitate is formed. After stirring for 30 minutes at -5 ° C., 1.50 g (8.51 mmol) of the compound obtained in step c / are added.

The mixture is then stirred at 0 ° C. for 40 minutes and then overnight at room temperature. 100 ml of dichloromethane are then added to this solution and then washed with 3 × 50 ml of 10% citric acid. After shaking vigorously, the phases are separated, the organic phase is washed with 2 × 50 ml of
NaHCO3 5%. After drying over MgSO 4, the solvent is removed in vacuo and the resulting crude product (3.00 g) is purified by recrystallization from CCl 4 (64% yield).

Mp 196-197 ° C
1 H-NMR (400 MHz, CDCl 3, ppm): 8.12 (1H, d, NH); 7.456.95 (9H, m, ArH); 5.30 (1H, d, NH); 5.25-5.0 (2H, m,
CH2Ph); 4.82 (1H, t, -H); 4.14 (1H, m, -H), 3.60-3.45 (2H, m, CH2indoline), 2.80 (3H, d, CH3-methylamide), 2.0-1.55 (2H, m.p. CH 2 aminobutyryl); 1.03 (1H, t, CH 3 aminobutyryl).

 Mass Spectrum (FAB): m / e (E intensity) = 396 (MH +, 9.9); 91 (C6Hs +, 100).

e / Preparation of the title compound as oxalate hydrate
Dissolved 2.15 g (5.44 mmol) of the compound obtained in step d / in 200 ml of methanol and 0.85 g of 10% palladium on activated charcoal (wet). The reaction mixture is hydrogenated at 60 psi for 3 hours. The solution is filtered and 1.3 equivalents (0.64 g) of oxalic acid (7.07 mmol) are added and the solution is concentrated in vacuo. Ethanol and then ether (1:50 ratio) are added and a crystallized product is obtained. After filtration and washing with diethyl ether, the solid is dried under vacuum for 24 hours on P2O5 at 35 ° C. 0.98 g of desired product are obtained (yield 49%).

Mp = 138-139 ° C
1 H-NMR (400 MHz, CD 3 OD, ppm): 8.20 and 8.02 (1H, d, ArH) 7.36-7.05 (3H, m, ArH); 5.18-5.02 and 4.70-4.62 (1H, m, aHindoline); 5.01-4.94 and 4.28-4.22 (1H, m, α-Haminobutyryl) 3.92-3.00 (2H, m, CH2indoline) 2.78 and 2.73 and 2.33 ( 3H, s,
CH3amide); 2.26-2.12 and 2.12-2.86 (2H, m, CH 2 aminobutyryl) 1.16 and 1.07 and 1.04 (3H, t, CH 3 aminobutyryl).

 Mass Spectrum (FAB): m / e (% intensity); 262 (MH +, 43.0); 154 (100); 137 (65.1); 136 (81.6).

 HPLC purity (Kromasil C-18, water / methanol / trifluoroacetic acid 70: 30: 0.1): 99.99%.

 IR spectrum (KBr disc, cm-1): 3440 (wave, NH); 3289 (m, NH); 2969 (m, CH); 2935 (m, CH); 1668 (s, C = O).

Microanalysis: C14H19N302. (COOH) 2.H20
Found: C, 51.55%; H = 6.05%; N = 11.28%
Calculated: C = 52.03%; H, 6.28%; N = 11.38%
Example 25: Preparation of 1- (2 (S) -aminobutyryl) 2 (S) -indolinecarboxylic acid ethylamide (UCL 1415-H); lane 3 R1 = CH2CH3; R2 = H; n = 0 and m = 1; R3 =

(R6 = H); R4 = CO-NH-R5; R5 = CH2CH3
Preparation of N-benzyloxycarbonyl 2 (S) -indolinecarboxylic acid ethylamide
The procedure is as in Example 24b using ethylamine (gas). The crude product obtained is purified by recrystallization from CCl4. Filtration afforded 4.05 g of pure amide as a white solid (74% yield).

PF = 201.0-201.5 ° C
Rf = 0.58 (ethyl acetate)
1 H-NMR (400 MHz, CDCl 3, ppm): 7.70 (1H, brs, COOH) 7.45-6.98 (9H, m, ArH); 5.90 (1H, brs, NH); 5.35-5.18 (2H, m, CH 2 benzyl); 4.90 (1H, dd, alpha -H); 3.64-3.04 (4H, m,
CH2indolinyl and NH-CH2); 0.99 (3H, brs, CH3).

b / Preparation of 2 (S) -indolinecarboxylic acid ethylamide
The procedure is as in Example 24 c / from the compound obtained in step a /. The resulting oil crystallizes after drying over P2O5 overnight. 2.25 g of desired product are obtained (95% yield).

PF = 92-95c
Rf = 0.28 (ethyl acetate)
1 H-NMR (400 MHz, ppm) 7.22-7.00 (3H, m, NH
ArH); 6.82 (1H, t, ArH); 6.73 (1H, d, ArH); 4.42 (1H, t, cx-H); 4.14 (1H, brs, NH); 3.65-3.50 (1H, m,
CH2indoline); 3.45-3.20 (2H, m, CH 2 CH 3); 3.14-3.02 (1H, m,
CH2indoline); 1.15 (3H, t, CH 3).

c / Preparation of 1- (N-benzyloxycarbonyl-2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid ethylamide
The procedure is as in Example 24 d / using acid
N-benzyloxycarbonyl-2 (S) -aminobutyric acid prepared as in Example 3 a / and the compound obtained from step b /. 2.23 g of pure amide are obtained in the form of a white solid (yield 50.5%).

Mp = 184.5-185 ° C 1 H-NMR (400 MHz, CDCl 3, ppm): 8.15 (1H, d, H aromatics)
J = 7.9 Hz); 7.44-7.08 (8H, m, ArH); 7.04 (s large,
NHamide); 5.37 (1H, d, NH); 5.0-5.2 (2H, m, PhCH 2) 4.84 (1H, m, α-H); 4.18 (1H, m, -H), 3.62-3.48 (2H, m, CH2indoline) 3.34-3.14 (2H, m, CH2ethylamide) 2 2.10-1, 60 (2H, m, CH 2 aminobutyryl); 1.20-0.98 (3H, m, CH 3).

d / Preparation of the title compound
The procedure is as in Example 24 c / for the compound from step c /. After drying the solid over P2O5 for 24 hours at 35 ° C., 1.51 g of desired product is obtained (yield 72%).

Mp = 115-120 ° C
1 H-NMR (400 MHz, D 2 O, ppm): 7.06-8.08 (4H, m, ArCH) 5.12 (1H, m, crH); 3.91 (1H, m, alphaH); 3.78-3.50 (1H, m, CH2Nindoline); 3.25-2.96 (3H, m, CH2Nindoline and NH-CH2) 2.25-1.90 (2H, m, CH2aminobutyryl); 1.28-0.98 (m,
CH3aminobutyryl and CH3ethylamide).

 Mass Spectrum (FAB). m / e (% intensity): 276 (MH +, 49.9); 191 (80.8); 118 (100); 89 (16.0) 77 (37.2); 58 (74.1); 51 (21.3); 50 (22.7); 46 (19.9); 39 (28.2); 31 (29.5).

 HPLC purity (Kromasil C-18, water / methanol / trifluoroacetic acid 65: 35: 0.1): 99.99%.

IR spectrum (Disc KBr, cm-1): 3395 (m, amide); 3057 (m,
NH3 +); 1718 and 1669 (s, C = O).

Microanalysis; C15H21N3O2. (COOH) 2.H2O
Found: C, 53.09%; H, 6.54%; N = 10.84%
Calculated: C, 53.26 z; H, 6.57%; N = 10.96%
Example 26: Preparation of 1-12 (S) -aminobutyryl) -2 (S) - (hydroxymethyl) indoline (UCL 1423-H); way 3.

R1 = CH2CH3; R2 = H; n = 0 and m = 1; R3 = (R6 = H); R4 = CH2OH

 The procedure is as for the previous examples according to route 3. The title compound is obtained in the form of an oxalate salt.

PF = 93-98 C
IR (KEr disc, cm-1): 3400 (NH3 +); 3160 (OH); 2975
(CH); 1652 (C = O); 1595 (C = C);
1 H-NMR (400 MHz, CD 3 OD, ppm): 8.06 (1H, d, aromatic) 7.29 (1H, d, aromatic H); 7.22 (1H, t, aromatic H); 7.09 (1H, t, aromatic H); 7.00 (4H, s, OH and NH3 +); 4.57 (1H, q, &alpha;H); 4.41 (1H, t, aH); 2.44 (3H, m, CH 2 indoline and 1H of
CH2O); 2.60 (1H, d, 1H CH2O); 1.97-1.75 (2H, m, CH 2 aminobutyryl); 0.97 (3H, t, CH 3 aminobutyryl).

Microanalysis: C13H18N202.C2H204.0.4H20
Found: C, 54.60%; H, 6.18%; N = 8.08%
Calculated: C, 54.34%; H, 6.32%; N = 8.45%
Example 27: Preparation of 1- (2 (S) -aminobutyryl) 2 (S) -indolinylmethyl-N-butylamine (UCL 1446-A2); lane 10
R1 = H; R2 = CH2CH3; n = 0 and m = 1; R3 =

(R6 = H); R4 = CH2-NH-RS; R5 = n.C4Hg
a / Preparation of N-CN-benzyloxycarbonyl-2 (S) -indolinylmethyl] butylamine: compound XXXII
To a solution (1M) of diborane in tetrahydrofuran at -78 ° C under nitrogen, N-benzyloxycarbonyl2 (S) -indolinylbutylamide (1.9 g, 5.6 mmol), prepared as in Example 22a, was added. in anhydrous tretrahydrofuran (25 ml). The mixture is heated and refluxed for 2 hours 30 minutes then cooled and methanol (11 ml) is added carefully. The mixture is stirred overnight and the solvent removed. The residue is dissolved in methanol (30 ml) and then evaporated. By column chromatography on silica gel, eluting with a methanol / chloroform mixture (5:95), 0.28 g of desired product is obtained (yield 16%).

PF = 36-38 ° C
1 H-NMR (CDCl 3, ppm): 7.48 (5H, m, PhCH); 6.96-7.13 (4H, dd, ArCH indoline); 5.25 (2H, s, PhCH 2); 4.54 (1H, brs, NCH indoline); 2.53-3.29 (4H, m, 2 x NCH 2); 1.30 (6H, m, 3 x CH 2); 0.85 (3H, t, CH 3).

Mass Spectrum (EI): m / e = 338 (M +)
Microanalysis: C21H26N202
Found: C, 74.44%; H = 7.90%
Calculated: C, 74.53%; H, 7.74%; N = 8.28%
b / Preparation of N- [N-benzyloxycarbonyl-2 (S) -indolinylmethyl] -N- (tert.butoxycarbonyl) butylamine: compound XXXIII
The compound from step a / (0.24 g, 0.7 mmol) is dissolved in aqueous dioxane (18 ml). 0.07 ml (0.7 mmol) of triethylamine and 0.35 g (1.4 mmol) of 2 (tert.butoxycarbonyloxyimino) -2-phenylacetonitrile are added. The mixture is stirred at room temperature for 18 hours and the solvent removed. The residue is purified by chromatography on silica gel (eluent chloroform / ethyl acetate 95: 5). 0.28 g is obtained in the form of an oil (yield 90%).

1 H-NMR (CDCl 3, ppm): 7.37 (5H, m, Ph); 7.07 (4H, m, ArCH indoline); 5.25 (2H, brs, PhCH2); 4.67 (1H, m, NCH) 3.14 (6H, m, 2 x NCH 2, ArCH 2 indoline); 1.34 (9H, s, tert.butoxycarbonyl); 1.15 (4H, m, CH 3 CH 2 CH 2); 0.81 (3H, t,
CH3).

 Mass Spectrum (EI): 438 (M +).

 c / Preparation of 2 (S) -tN-butyl-N- (tert.butoxycarbonyl) aminomethyl] indoline: compound XXXIV.

 The compound obtained in the preceding step (0.28 g, 1 mmol) is hydrogenated in aqueous methanol medium (10 ml) containing palladium on activated carbon at 10% (0.11 g) for 3 hours 30 minutes. After removal of the catalyst and concentration of the solution, 0.19 g (100% yield) of the product is obtained.

1 H-NMR (CDCl 3, ppm): 6.73-7.04 (4H, m, ArCH); 4.14 (1H, brs, NH); 4.02 (1H, m, NHCH); 3.25 (2H, m, NCH 2); 2.93
(2H, d, NCH2); 1.18-1.68 (6H, m, 3 x CH 2); 1.38 (9H, s, tert.butoxycarbonyl); 0.88 (3H, t, CH 3).

 Mass spectrum (EI): m / e = 304 (M +).

d / Preparation of 2- [N- (tert.butoxycarbonyl) -N-butylamino-methyl) -N-EN- (tert.butyoxycarbonyl) -2 (S) -aminobutyryl] indoline: Compound XXXV
To the compound obtained by Example 2a / (0.6 g, 2.95 mmol), dissolved in dimethylformamide (20 ml) and dichloromethane (20 ml), cooled to -5 ° C., 0.12 g are added ( 0.58 mmol) of dicyclohexylcarbodiimide and the mixture is stirred for 5 minutes, then 91 mg (0.56 mmol) of 3-hydroxy-1,2,3-benzotriazine-4 (3H) -one is added. The mixture is stirred at 0 ° C. for one hour and then 170 mg (0.56 mmol) of the compound obtained by step c / are added. The mixture is stirred at room temperature overnight, the solvent removed and the residue purified on a silica gel chromatography column (eluent ethyl acetate / petroleum 4: 1). 90 mg of product is obtained (yield 33%) .

1 H-NMR (CDCl 3, ppm): 8.16 (1H, brs, ArCH); 7.12 (3H, m, ArCH); 4.56 (1H, m, NCHCO); 4.25 (1H, m, NCHCH2N); 3.143.40 (4H, m, 2 x NCH 2); 1.12-1.81 (8H, m, 4 x CH 2); 1.34
(18H, s, 2 x tert.butoxycarbonyl); 0.8-1.0 (6H, m, 2 x CH 3).

 Mass spectrum (EI): m / e = 489 (M +).

e / Preparation of the title compound as dihydrochloride
The compound obtained in the preceding step (90 mg, 0.184 mmol) is treated with a cold mixture of concentrated HCl (1 ml) and glacial acetic acid (9 ml). The solution is stirred for 1 hour 30 minutes and the solvents removed to obtain the product.

 Mp = 113-8 ° C.

 HPLC purity (Kromasil C-18, water / methanol / trifluoroacetic acid 70: 30: 0.1). 100%.

 1 H-NMR (CD 30 D, ppm) 8.01 (1H, m, ArCH) 7.0-7.28 (3H, m, ArCH); 4.31 (1H, m, NCHCO); 3.91 (1H, t, NCHCH2N); 2.953.08 (4H, m, 2 x CH2); 1.88 (2H, m, CH 2); 1.64 (2H, m, CH 2) 1.32 (2H, dd, CH 2); 1.08 (2H, m, CH 2); 0.92 (3H, t, CH 3) 0.86 (3H, t, CH 3).

Mass Spectrum (FAB): 290 (M + + 1)
Microanalysis: C17H29N3OCl2.1,8H2O:
Found: C, 51.83%; HH = 8.21%; N = 10.34%;
Calculated: C, 51.72%; H, 8.32%; N = 10.64%.

Example 28 Preparation of 1- (2 (S) -aminobutyryl) 2 (R / S) - (5-methoxy) indolinecarboxylic acid n-butylamide (UCL 1486
H); lane 12
R1 = CH2CH3; R2 = H; n = 0 and m = 1; R3 =

R6 = OCH3; R4 = CO-NH-R5; R5 = n. C4 H9
a / Preparation of the ethyl ester of 5-methoxy2-indolecarboxylic acid: compound XXXXII
P-Anisidine (61.5 g, 499 mmol) is treated in 500 ml of 15% HCl containing 500 g of ice by adding dropwise for 5 minutes a solution of sodium nitrite (37.5 g). , 543 mmol) in water (100 ml). 3 g of activated charcoal are added and the mixture is stirred for 10 minutes. After filtration, the filtrate is rapidly added to ethyl 2-ethylacetoacetate (80 g, 554 mmol) and anhydrous sodium acetate (410 g, 4.998 mol). ), in ethanol (500 ml) containing 500 g of ice. The mixture is stirred for 2 hours and then extracted with toluene (3 × 500 ml). The extracts are dried (Na2SO4) and the solvent removed to obtain crude ethyl 1- (p-methoxyphenylazo) -1-methyl acetoacetate as an oil. This oil is added to a cold solution of HCl in methanol (38-40% by weight). The mixture is heated and refluxed for 40 minutes. The mixture is stirred for 2 hours at room temperature, added with water (150 ml) and then refrigerated. The product separates in the form of orange crystals which are collected and washed with cold methanol and then with water. 79.5 g (yield 72.6%) are obtained.

 1 H-NMR (CDCl 3, ppm): 9.03 (1H, brs, NH); 7.31 (1H, dd, ArCH); 7.14 (1H, t, ArCH); 7.07 (1H, d, ArCH); 6.99 (1H, dd, ArCH); 4.40 (2H, q, CH 2); 3.84 (3H, s, CH 3 O); 1.43 (3H, t, CH 3 CH 2).

 b / Preparation of 5-methoxy-2 (R / S) -indoline carboxylic acid n-butylamide: compound XXXXIV.

The product from step a / (10 g, 53 mmol) is dissolved in methanol (100 ml) and treated with magnesium (3.6 g, 150 mmol). The reaction temperature is maintained at 15-20 ° C by external cooling and the mixture stirred overnight, then diluted with 300 ml of water and extracted with methylene chloride (4 x 100 ml). dried (Na2SO4) and the solvent removed to give 5-methoxy-2 (R / S) -indolinecarboxylic acid methyl ester (compound
XXXXIII). This is dissolved in butylamine (40 ml) and refluxed under nitrogen for 3 hours. Excess amine was removed and the residue crystallized from ether to give XXXXIV (9.00 g, 82% yield).

 Mp 119-121 ° C.

1 H-NMR (CDCl 3, ppm): 7.2 (1H, s, NHCO); 6.6-6.7 (3H, m,
ArCH); 4.37 (1H, t, NHCHCO); 3.90 (1H, s, NH indoline); 3.73 (3H, s, OCH3); 3.54 (1H, dd, ArC (H) H); 3.27 (2H, m, NHCH 2) 3.05 (1H, dd, Ar-C (H) H); 1.2-1.5 (4H, m, CH 3 (CH 2) 2), 0.89 (3H, t, CH 3 (CH 2) 2) -
c / Preparation of 1 - (N-benzyloxycarbonyl) -2 (S) -aminobutyryl) -2 (R / S) - (5-methoxy) indolinecarboxylic acid n-butylamide: compound XXXXV
2.37 g (10 mmol) of N-benzyloxycarbonyl-2 (S) -aminobutyric acid, obtained according to Example 3a /, and the compound obtained by step b / (2.48 g, 10 mmol) are dissolved in anhydrous methylene chloride and cooled to 0 ° C. The mixture is treated with triethylamine (2.03 g, 20 mmol) and then with bis (2-oxo-3-oxazolidinyl) phosphinic chloride (2.8 g 10 mmol). The reaction mixture is stirred overnight at room temperature. It is washed with a 96% citric acid solution (3 x 100 ml) then a 5% sodium bicarbonate solution (3 x 100 ml), dried (Na 2 SO 4) and the solvent is removed. The product is purified by chromatography on silica gel, eluent ethyl acetate / petroleum 1: 2. 3.4 g (yield 72.6%) are obtained.

1 H-NMR (CDCl 3, ppm): 8.0 (0.5H, m, NH); 7.2-7.3 (5H, m,
Ph); 6.6-7.1 (3.5H, m, NH and ArCH indoline); 4.7-5.7 (4H, m,
PhCH2, NH, NCHCO of butyric acid); 4.0-4.3 (1H, m, NCHCO indoline); 3.78 (3H, s, OCH3); 3.45 (1H, m, ArCH (H) indoline); 3.0-3.2 (3H, m, NHCH2, ArCH (H) indoline); 0.7-2.0 (12H, 3 x CH2, 2 x CH3).

 Mass spectrum (FAB): m / e = 468 (M + + 1).

d / Preparation of the title compound in the form of oxalate salt
The compound of step c / (2.8 g, 6 mmol) is dissolved in 100 ml of palladium-on-charcoal-containing methanol (10%, 0.8 g, 50% moisture) and the mixture is hydrogenated at room temperature. 40 psi for 2.5 hours. The catalyst is removed by filtration and the solvent removed. The residue is treated with oxalic acid (0.85 g, 9.4 mmol) in ethanol (10 ml) and then ether (500 ml). The product precipitates. 1.86 g (72% yield) are obtained.

Microanalysis: C18H27N303. (COOH) 2.0,25H20
Found: C = 56, 14%; H, 6.98%; N = 9.57%
Calculated: C, 56.13%; H = 6.95%; N = 9.82%
Mass Spectrum (FAB): m / e = 333 (M +)
1 H-NMR (DMSO-d6, ppm): 8.3-9.1 (concealed ArCH); 7.97 (1H, dd, CONH); 6.7-7.0 (2H, m, ArCH); 4.9-5.3 (1H, m, NCHCO indoline); 3.2-3.9 (5H, m, CH 3 O, NCHCO butyl, ArCH (H)); 2.73.2 (3H, m, CONHCH2, ArCH (H)); 1.0-2.0 (6H, m, CH 3 CH 2,
CH3 (CH2) 2); 0.7-1.0 (2 x CH3CH2).

 HPLC purity (Kromasil C-18 J Methanol / water / trifluoroacetic acid 50: 50: 0.1): 99.92%.

(Diastereoisomeric ratio 39.37: 60.55)
e / Optical separation of 1- (2 (S) -aminobutyryl) 2 (R / S) - (5-methoxy) indolinecarboxylic acid n-butylamide acid (UCL 1486
H)
1- (N-Benzyloxycarbonyl-2 (S) -aminobutyryl 2 (R / S) - (5-methoxy) indolinecarboxylic acid n-butylamide
XXXXV) is separated into its diastereoisomers by column chromatography on silica gel using a mixture of methylene chloride / diethyl ether (10: 1) as eluent.

 The (S, S) isomer is eluted first. It is collected and recrystallized from methanol and then the benzyloxycarbonyl protecting group is removed by hydrogenation in the presence of palladium on charcoal in methanol for 2 hours and 30 minutes under a hydrogen pressure of 60 psi, as described above. The catalyst is removed by filtration and the residue is treated with oxalic acid in methanol and then ether to yield the oxalate salt (UCL 1704-H).

Mp = 185-190 ° C
Microanalysis: C18H27N3O3.1,4C2H2O4.0,5H2O:
Found: C, 53.3%; H, 6.81%; N = 9.05%
Calculated: C, 53.3%; H, 6.63%; N = 8.97%
The isomer (S, R) is eluted second. It is treated in a manner similar to the (S, S) isomer to give the oxalate salt (UCL 1705-H).

PF = 95-105 ° C
Microanalysis: C18H27N303. 1, 35C2H204
Found: C, 54.4%; H = 6.55%; N = 9.45%
Calculated: C, 54.1%; H, 6.58%; N = 9.24%
Example 29 Preparation of 1- (2 (S) -aminobutyryl) 2 (R / S) - (6-methoxy) indolinecarboxylic acid n-butylamide (UCL 1546
H); lane 12
R1 = CH2CH3; R2 = H; n = 0 and m = 1; R3 =

R6 = OCH3; R4 = CO-NH-R5; R5 = n.C4Hg
The procedure is as in Example 28. The title compound is obtained in the form of an oxalate salt with a yield of 70%.

Microanalysis: C18H27N303. (COOH) 2.H20
Found: C, 54.54%; HH = 7.08%; N = 9.52%
Calculated: C, 54.41%; H = 7.0896; N, 9.52%;
1 H-NMR (DMSO-d6, ppm): 8.5 (1H, b, NH); 7.3-8.2 (6H, m,
NH3 +, ArCH, H2O); 7.0-7.2 (1H, m, ArCH); 6.5-6.7 (1H, m,
ArCH); 5.05 (1H, dd, NCHCOindoline); 3.2-4.1 (5H, m, CH 3 O, ArCR (H), NCHCO butyryl); 2.7-3.2 (3H, m, NCH2, ArCH (H)) 1.5-2.0 (2H, m, CH2); 1.0-1.5 (4H, m, 2 x CH2); 0.7-1.0 (6H, m, 2 x CH3).

 HPLC purity (Kromasil C-18, methanol / water / trifluoroacetic acid 50: 50: 0.1): 98.2%.

(Diastereoisomeric ratio 39.4: 58.8)
Example 30: Preparation of 1- (2 (S) -aminobutyryl) 2 (R / S) - (5-fluoro) indolinecarboxylic acid n-butylamide (UCL 1534
H); lane 12 R1 = CH2CH3; R2 = H; n = 0 and m = 1; R3 =

R6 = F; R4 = CO-NH-R5; R5 = n.C4H9
a / Preparation of the ethyl ester of (5-fluoro) 2-indole carboxylic acid: compound XXXXII
The procedure is as in Example 28a / to prepare the ethyl 1- [p-fluorophenylazo) -1-methyl acetoacetate from p-fluoroaniiine and ethyl 2-methylacetoacetate (crude yield 62g). It is dissolved in 270 ml of acetic acid containing 15 ml of sulfuric acid, and then the mixture is refluxed for 15 minutes. After cooling to room temperature, 500 ml of water are added and the mixture is extracted with water. ether (500 ml) and methylene chloride (500 ml). The combined extracts are washed with water (1000 ml) and with 10% potassium carbonate solution (300 ml), dried and the solvent removed. The product is then obtained by crystallization in ethanol. 6.8 g are obtained (yield 14%).

1 H-NMR (CDCl 3, ppm): 7.0-7.4 (4H, m, ArCH); 4.41 (2H, q,
CH2); 1.40 (3H, t, CH 3).

 b / Preparation of 5-fluoro-2 (R / S) -indoline carboxylic acid n-butylamide: compound XXXXIV.

 The procedure is as in Example 28BI. Yield 62 9s.

 Mp 93-50C.

1 H NMR CDCl 3, ppm) 7.12 (1H, b, NH); 6.6-6.9 (3H, m,
ArCH); 4.42 (1H, t, NHCHCO); 4.03 (1H, b, NH); 3.56 (1H, dd,
ArCH (H)); 3.27 (2H, m, CH 2 NH); 3.05 (1H, m, ArCH (H)); 1,2- 1,7 (CH3 (CH2) 2); 0.90 (3H, t, CH 3).

 Mass spectrum (FAB): m / e = 237 (M + + 1).

c / Preparation of 1- (N- (benzyloxycarbonyl) -2 (S) -aminobutyryl) -2 (R / S) - (5-fluoro) indolinecarboxylic acid n-butylamide: compound XXXXV
The procedure is as in Example 28c / and then the product is purified by chromatography on silica gel (eluent ethyl acetate / petroleum 1: 1). Yield 54R.

 M.p. 192-3 C.

1 H-NMR (CDCl 3, ppm): 8.05 (1H, b, NH); 7.2-7.4 (5H, m,
Ph); 6.8-7.1 (4H, m, NH, ArCH indoline); 4.7-5.4 (4H, m,
PhCH2, 2 x NCHCO); 3.5 (2H, m, ArCH 2 indoline); 3.2 (2H, m,
CH2NHCO); 0.7-2.0 (12H, m, CH3CH2 and CH3 (CH2) 2).

dl Preparation of the title compound as the oxalate salt
The procedure is as in Example 28 d /. Yield 68%.

Microanalysis: C17H24FN302. (COOH) 2.0,75H20
Found: C, 53.58%; H, 6.60%; N = 10.25%
Calculated: C, 53.70%; H = 6.52%; N = 9.89%
1 H-NMR (DMSO-d6, ppm): 8.60 (1H, b, NH); 7.8-8.1 (1H, m,
ArCH); 7.2 -7.8 (4H, b, NH3 + and H2O); 6.8-7.2 (2H, m, ArCH) 5.0-5.2 (1H, m, NHCHCO indoline); 2.6-3.8 (5H, m, NCHCO butyryl, ArCH2, CH2NHCO); 1.5-2.0 (2H, m, CH 2); 1.0-1.5 (4H, m, 2 x CH2); 0.6-1.0 (6H, m, 2 x CH3).

 Mass Spectrum (FAB): m / e = 322 (M ++ 1).

 HPLC purity (Kromasil C-18, methanol / water / trifluoroacetic acid 50: 50: 0.1): 100%.

(Diastereomeric ratio 36.1: 63.9)
Optical separation of 1- (2 (S) -aminobutyryl) 2 (R / S) - (5-fluoro) indolinecarboxylic acid n-butylamide (UCL 1534-H)
1- (N- (benzyloxycarbonyl) -2 (S) -amino-butyryl) -2 (R / S) - (5-fluoro) indolinecarboxylic acid is separated into its diastereoisomers by column chromatography on silica gel using as eluent a mixture of petroleum ether
(PE = 60-80 ° C) / ethyl ethanoate (2: 1).

The isomer (S, R) is eluted first. It is recrystallized from a petroleum / diethyl ether mixture and then deprotected by hydrogenation as indicated in Example 28 el. We obtain
the (S, R) isomer in the form of oxalate salt (UCL 1703-H).

Mp 173-177 ° C
Analysis: C17H24FN3O2 .C2H204.H2O
Found: C, 52.9%; H, 6.37%; N = 9.52%
Calculated: C, 53.1%; H, 6.57%; N = 9.78%
The (S, S) isomer is eluted second and treated in a manner analogous to the first isomer for purification and deprotection. The isomer (S, S) is also obtained in the form of oxalate salt (UCL 1702-H).

Mp 105-110 ° C
Microanalysis: C17H24FN302.C2H204.H20
Found: C, 52.9%; H, 6.37%; N = 9.70%
Calculated: C, 53.1%; H, 6.57%; N = 9.78%
Example 31: Preparation of 1- (2 (S) -aminobutyryl) 2 (R / S) - (5-benzyloxy) indolinecarboxylic acid n-butylamide (UCL 1535-A); lane 12 R1 = CH2CH3; R2 = H; n = 0 and m = 1; R3 =

R6 = OCH2Ph; R4 = CO-NH-R5; R5 = n.C4Hg
The procedure is as in Example 28.

1- (N- (tert.butoxycarbonyl) -2 (S) -aminobutyryl) -2 (R / S) - (5-benzyloxy) indolinecarboxylic acid n-butylamide (3.56 g, 7 mmol) is dissolved in ethyl acetate (20 ml) then treated with HCl in ethyl acetate (3M, 25 ml). The mixture is stirred at room temperature for 1 h and the solid formed is collected, washed and dried. The title compound is isolated as the hydrochloride salt. 0.9 g is obtained
(Yield 30.2%).

Microanalysis: C24H32N303C1
Found: C, 63.28%; H = 6.92%; N = 9.20%
Calculated: C, 63.36%; H, 7.31%; N = 9.24%
1 H-NMR (DMSO-d6, ppm). 8.7 (0.9H, brs, NH of a diastereoisomer); 8.5 (brs, NH3 + and H2O); 8.07 (0.1H, NH of the second diastereoisomer); 7.93 (1H, d, ArCH); 7.2-7.5 (5H, m, ArCH); 6.93 (1H, s, ArCH); 6.84 (1H, dd, ArCH); 5.35
(0.9H, dd, NCHCO indoline of a diastereoisomer); 4.9-5.2 (2.1H, PhCH2, NCHCO indoline of the second diastereoisomer); 4.7 (0.1H, m, NCHCO butyryl of a diastereoisomer); 3.92 (0.9H, m, NCHCO butyryl of the second diastereoisomer); 3.4-3.7 (1H, m,
ArCH (H) indoline); 2.8-3.1 (3H, m, ArCH (H) indoline, NCH 2) 1.5-2.0 (2H, m, CH 2); 1.05-1.5 (4H, m, 2 x CH2); 0.7-1.0 (6H, m, 2 x CH3).

 Mass spectrum (FAB): m / e = 410 (M ++ 1).

 HPLC purity (Kromasil C-18, methanol / water / trifluoroacetic acid 70: 30: 0.1): 98.93 *.

 (Diastereomeric ratio 85.83: 13.1).

Example 32: Preparation of 1- (2 (S) -aminobutyryl) 2 (S) -t (3aS, 7aS) -perhydro] indolinecarboxylic acid n-butylamide (UCL 1442-H); lane 3 R1 = CH2CH3; R2 = H; n = 0 and m = 1; R3 =

R4 = CO-NH-R5; R5 = n.C4Hg
a / Preparation of 2 (S) - [(3aS, 7aS) perhydro] indoline carboxylic acid as N-acetate salt.

 A solution of 2 (S) -indolinecarboxylic acid (2.5 g, 15.32 mmol) in acetic acid containing PtO 2 (1 g) is hydrogenated at 55 ° C for 2 hours at 60 psi. The catalyst is removed by filtration and the solvent removed in vacuo to give a crude residue which is crystallized from diethyl ether to give, after filtration and washing with ether, a white solid. 1.28 g are obtained (yield 27.5%).

 M.p. 247-248 C.

1 H-NMR (200 MHz, CD 3 OD, ppm) 4.05-3.90 (1H, m, Hunction); 3.70-3.55 (1H, m, alphaH); 2.45 - 2.20 (2H, m, CH 2
COOH); 2.15-1.25 (12H, m, 1-cyclohexyl, H junction and CH 3
AcOH).

 b / Preparation of N- (benzyloxycarbonyl) -2 (S) -t (3aS, 7aS) perhydro] indolinecarboxylic acid.

 A solution of the compound obtained in step a / (1.28 g, 5.58 mmol) in 2N NaOH (30 ml) is cooled in an ice-water bath and stirred vigorously. Benzyl chloroformate (0.95 mL, 6.65 mmol) and 2N sodium hydroxide (30 mL) are added (in about 10 parts). The temperature of the reaction mixture is maintained between 5 and 10 C by adjusting the rate of addition of the reagents. 40 ml of 2N sodium hydroxide are added and the mixture is kept at room temperature for 30 minutes. The alkaline solution is washed with ether (4 x 50 ml) and then acidified to pH <2 by addition of 5N HCl. The solution is extracted with CH 2 Cl 2 and the extracts are dried over MgSO 4, filtered and evaporated under vacuum. The oil obtained is dried under vacuum over P 2 O 5 to obtain 1.40 g of the desired product (yield 83.5%).

 1 H-NMR (200 MHz, CDCl 3, ppm): 7.14 (5H, brs, ArCH) 5.05 (2H, brs, benzyl H); 4.20 - 4.10 (1H, m, Hunction) 4.00-3.75 (1H, m, AH); 2.20-1.00 (11H, m, CH 2 cyclohexyl, H, CH 2 COOH).

cl Preparation of N-benzyloxycarbonyl2 (S) - [(3aS, 7aS) perhydro] indolinecarboxylic acid n.butylamide
The procedure is as in Example 15 step c / using the product obtained in Example 32 step b /.

1.40 g of the product from the previous step are dissolved
(4.61 mmol) in 50 ml of anhydrous tetrahydrofuran and 0.58 ml of 4-ethylmorpholine (4.61 mmol) are added at a temperature of 0 ° C. This solution is cooled to -5 ° C. and 0.60 ml. Isobutylchloroformate (4.61 mmol) is added dropwise. After 30 minutes at -5 ° C., 0.46 ml of butylamine (4.61 mmol) are added to the solution and then the mixture is left overnight at room temperature. 70 ml of dichloro-methane are then added and this solution is washed with 3x50 ml of 10% citric acid. After vigorous stirring, the phases are separated. The organic phase is washed with 2 × 50 ml of 5% NaHCO 3. After drying over MgSO 4, the solution is evaporated under vacuum and 1.70 g of crude amide (yield 98%) is obtained which is used without further purification.

1 H-NMR (200 MHz, CDCl 3, ppm): 7.30 (5H, brs, ArCH) 5.05 (2H, brs, benzyl H); 4.20 (1H, m, oeH); 4.00-3.80 (1H, m, H junction); 3.80-3.60 (1H, m, NH); 3.20-3.00 (2H, m,
CH2-NH butylamide); 2.30-1.00 (15H, m, CH 2 cyclohexyl, CH 2
CH2-BuNH, Hjunction, CH2-COOH); 1.00-0.80 (6H, m, butylamide CH3).

d / Preparation of 2 - [(2S, 3aS, 7aS) perhydro] indoline carboxylic acid n. butylamide
The product of step cJ (1.70 g, 4.54 mmol) in solution in 200 ml of methanol is hydrogenated at room temperature for 2 hours 30 minutes at 60 psi using as catalyst 0.6 g of Pd / C (10%, wet). The amide is isolated as an oil.

 1 H-NMR (200 MHz, CDCl 3, ppm): 7.80 (1H, brs, NH) 3.90-3.80 (1H, m, crH); 3.50-3.00 (3H, m, -CH 2 -NH-BuNH-, H junction); 3.00-1.05 (15H, m, -CH 2 -cyclohexyl, H, -CH 2 -butylamide); 0.95 (3H, t, butylamide CH3).

 e / Preparation of 1- (N-benzyloxycarbonyl) -2 (S) aminobutyryl) -2 (S) - [(3aS, 7aS) perhydro] indolinecarboxylic acid n -butylamide.

N-benzyloxycarbonyl-2 (S) aminobutyric acid obtained as in Example 3 was dissolved in 45 ml of anhydrous tetrahydrofuran and 0.51 ml of 4-ethylmorpholine was added (0.95 g, 4 mmol). (4.01 mmol) at 0 ° C. This solution is cooled to -5 ° C. and treated drop by drop with
isobutylchloroformate (0.52 ml, 4.01 mmol). After 30 minutes at -5 ° C., the compound of step d is added to the mixture.
(0.90 g, 4.01 mmol) in solution in 5 ml of anhydrous tetrahydrofuran and the mixture thus obtained is left overnight at room temperature. 50 ml of dichloromethane are then added, and then the mixture is washed with 3 × 50 ml of a 10% citric acid solution. The organic phase is
washed with 2 x 50 ml of 5% NaHCO 3. After drying over MgSO 4, the solution is evaporated under vacuum and a crude product is obtained which is purified by flash chromatography on silica eluting with an ethyl acetate / ether mixture (15:85 then 35:65). This makes it possible to obtain 0.66 g of the desired product in the form of an oil. It is dried overnight under reduced pressure on P @ O @.

 1 H-NMR (200 MHz, CDCl 3, ppm): 7.20 (5H, brs, ArCH) 6.90 (1H, brs, -NH-); 5.45 (1H, d, -NH-); 5.10-4.95 (2H, m, H benzyl); 4.50-4.30 (2H, m, H junction and aH); 4.10-4.00 (1H, m, c * H); 3.20-3.00 (2H, m, -CH 2 -butylamide); 2.65-1.00 (17H, m, -CH 2 -cyclohexyl, H, -CH 2 -aminobutyryl, -CH 2 -butylamide); 1.00-0.90 (6H, m, CH 3 aminobutyryl and CH 3 butylamide).

 Mass Spectrum (FAB) m / e (% Intensity): 444 (MH +, 13.5); 225 (56); 124 (100); 91 (91).

f / Preparation of the title compound
A solution of the compound obtained in step e / (0.66 g, 1.49 mmol) in 100 ml of methanol is hydrogenated for 3 hours using a Pd / C catalyst (0.25 g, 10%, wet). at 60 Psi.

The catalyst is removed by filtration and the solvent evaporated in vacuo. 0.18 g of oxalic acid (1.98 mmol) is added to the crude oil thus obtained in ethanol and then ether is added thereto. After crystallization, filtration and drying under vacuum, 0.375 g (62.5%) of the desired product is obtained.

PF: 168-9 C (open capillary)
1 H-NMR (400 MHz, CD30D, ppm): 4.30-4.25 (1H, m, aH) 4.05-3.85 and 3.40-3.35 (2H, m, aH and 7aH) ; 3.15-2.98 (2H, m, -CH 2 -NH-BuNH 2); 2.33-2.15 (1H, m, 3aH); 2.05-1.50 (9H, m, -CH 2 -aminobutyryl, -CH 2 -perhydroindoline); 1.45-1.05 (7H, m, -CH 2 -CH 2 -BuNH 2 and H proline); 0.95-0.75 (6H, m, -CH 3 aminobutyryl and -CH 3 BuNH 2).

HPLC (column: Kromasil C-18 (250 mm x 4.6) 5 pm detector: uv 215 nm; flow rate: 1 ml / minute; mobile phase
A / B = 50/50 with A = H20 + 0.1% trifluoroacetic acid and B =
CH 3 OH + 0.1% trifluoroacetic acid): majority peak 98.5%, retention time = 19.8 minutes; minority peak 1.4%, retention time = 18.9 minutes.

Mass Spectrum (FAB). m / e (% intensity) = 332 (MH +
Na) +, 1); 310 (MH +, 100); 225 (75); 124 (66); 59 (C7H7 +, 43).

 IR (KBr disc, cm1): 3342 (NH3 +, wave); 3049 (NH, wave); 2937 (CH, s); 2857 (CH, s); 1745 (C = O, s); 1682 (s); 1652 (s).

Microanalysis: C17H31N2O2.1.0C2H2O4.0.3H2O
Found: C = 56.34 *; H, 8.37%; N = 10.33%
Calculated: C, 56.36%; H, 8.36%; N = 10.38%
Example 33: Preparation of 2- (2 (S) -aminobutyryl) 1- (R / S) -isoindolinecarboxylic acid n-butylamide acid (UCL 1466-H); lane 4 R1 = CH2CH3; R2 = H; n = 1 and m = 0; R3 =

R4 = CO-NH-R5; R5 = n.C4Hg
al Preparation of the acid o. (Bromomethyl) phenylacetic acid.

O-tolylacetic acid (40 g, 266 mmol), N-bromosuccinimide (52.147 g, 293 mmol) and benzoyl peroxide (0.174 g, catalytic) are dissolved in 300 ml of CCl 3. The suspension is refluxed and kept there for 12 hours. After cooling, the solid particles are separated by filtration and the CCl 4 is evaporated in vacuo,
revealing a white solid. It is sufficiently pure without recrystallization (90% yield).

1 H-NMR (CDCl 3, ppm): 7.25 (4H, m, ArH); 4.3 (2H, s,
CH2COOH); 3.7 (2H, s, CH 2 Br).

 b / Preparation of the methyl ester of o. (bromomethyl) phenylbromoacetic acid.

 The acid from step a / is dissolved in 44 ml of BASE and heated for 3 hours at 55 ° C. The SOCL @ is removed under vacuum and the resulting brown oil is treated with 26 ml of Br: by irradiating with a 450 W UV lamp for 2 hours.

After addition, the bromine is removed by vacuum distillation and the black residue is poured into 305 ml of methanol. The methanol is then removed and there remains a black oil which is distilled under a pressure of 0.1 mm Hg. The distallation fractions leaving between 120 and 1300C are recovered (Yield 60%).

1 H-NMR (CDCl 3, ppm): 7.0-7.50 (4H, m, ArH); 5.7 (1H, s,
CH); 4.3 (2H, d, CH 2 Br); 3.7 (3H, s, CH3).

 cl Preparation of N-benzyl (R / S) -isoindoline carboxylic acid methyl ester as hydrochloride.

 The methyl ester (46.4 g, 144 mmol) from step b / is dissolved in anhydrous toluene (288 mL) under nitrogen. 46.35 g of benzylamine (864 ml) are added using an ice / salt mixture. The mixture is stirred for 72 hours. Then, the amine hydrobromide is removed by filtration and the toluene evacuated under vacuum. The yellow oil obtained is dissolved in ether and HCl is bubbled to obtain the hydrochloride (yield 50%).

PF: 160-1610C
1 H-NMR (D 2 O, ppm): 7.00 (9H, m, ArH); 5.20 (1H, s, aH) 4.10 (2H, d, CH 2 ring); 4.00 (2H, d, CH 2 Bn) 3 3.10 (3H, s,
CH3).

d / Preparation of the methyl ester of the acid 1 (R / S) isoindoline carboxylic in the form of hydrochloride
The product of step c / (5 g, 16 mmol) is dissolved in 100 ml of ethanol and Pd / C (10%, wet) is added. The suspension is stirred under 40 psi of hydrogen at 500C and for 8 hours. The catalyst is filtered off and the ethanol concentrated to a small volume. The addition of ether makes it possible to show white shoots. The degree of purity of the product is sufficient and does not require recrystallization (Yield 90%).

PF: 159-1610C
1 H-NMR (CD30D, ppm): 7.25-7.30 (4H, m, ArH); 5.50 (1H, s, H); 4.50 (2H, d, CH 2 cycle); 3.8 (3H, s, CH 3).

 e / Preparation of the methyl ester of 2- (N- (benzyloxycarbonyl) -2 (S) -aminobutyryl) -1 (R / S) -isoindole carboxylic acid.

The N-benzyloxycarbonyl-2 (S) -aminobutyric acid (compound of Example 3 step a /) (2.62 g, 11.04 mmol) is dissolved in 100 ml of tetrahydrofuran under nitrogen and cooled on an ice-cream mixture. /salt. N-ethylmorpholine (1.27 g, 11.04 mmol) is added followed by isobutyl chloroformate (1.51 g, 11.04 mmol). After 30 minutes, the compound obtained from step d / (2.347 g, 11.04 mmol) is added. Stirring is maintained overnight. The mixture is then poured into 200 ml of
CH 2 Cl 2 washed with 3 × 100 ml of citric acid (10%) and then with 3 × 100 ml of NaHCO 2 (58), dried and evaporated to a clear oil (50% yield).

 1 H-NMR (CDCl 3, ppm): 7.25 (9H, m, ArH); 5.75 (1H, m, csH); 5.10 (2H, s, CH 2 benzyloxycarbonyl); 4.60-5.00 (3H, m, H, CH 2 cycle); 3.8 (3H, s, OCH3); 1.60-2.00 (2H, m, CH 2 aminobutyryl); 1.0 (3H, m, CH 3 aminobutyryl).

 f. Preparation of 2- (N-benzyloxycarbonyl) (S) -aminobutyryl) -1 (R / S) -isoindolinecarboxylic acid.

 The compound of step e / (3.422 g, 8.6 mmol) is dissolved in 69 ml of methanol. 8.6 ml of 2N NaOH are added and the solution is stirred at room temperature for 24 hours. 172 ml of water are then added and the methanol is evacuated under vacuum. The solution is acidified with concentrated HCl to pH 2 and the resulting suspension is extracted with CHrCl2. After drying, the CH 2 Cl 2 is evacuated. It remains an oil (Yield 90%).

 1 H-NMR (CDCl 3, ppm): 7.00-7.30 (9H, m, ArH); 5.75 (2H, m, CH 2 cycle); 4.8-5.2 (3H, m, zH, CH 2 benzyloxycarbonyl) 4.60 (1H, m, alphaH); 1.90 (2H, m, CH 2 aminobutyryl); 1.0 (3H, m, CH 3 aminobutyuryl).

 g / Preparation of 2- (N-benzyloxycarbonyl) (S) -aminobutyryl) -1 (R / S) -isoindolinecarboxylic acid n-butylamide.

 The compound of step f1 (2.70 g, 7.07 mmol) is dissolved in 100 ml of tetrahydrofuran under nitrogen and cooled on an ice / salt mixture. N-ethylmorpholine (0.814 g, 7.07 mmol) is then added, followed by isobutylchloroformate (0.966 g, 7.07 mmol). After 30 minutes, n-butylamine (0.517 g, 7.07 mmol) was added and stirring was continued overnight. The mixture is poured into 200 ml of CH 2 Cl 2, washed with 3 × 100 ml of citric acid (108) and then with 3 × 100 ml of NaHCO 3 (5%), dried and evaporated until a clear oil is obtained (yield 80%). .

1 H-NMR (CDCl3, ppm) 7.25 (9H, m, ArH); 5.20-5.30 (2H, m, CH 2 cycle); 4.80-5.10 (3H, m, zH, CH 2 benzyloxycarbonyl) 4.00-4.25 (1H, m, aH); 3.0 (2H, m, CH 2 N); 1.00-1.90 (6H, m,
CH2 aminobutyryl, CH3CH2CH2); 0.8-1.5 (6H, m, CH 3 aminobutyryl, butyl CH 3).

h / Preparation of the title compound as the oxalate salt
The compound of step g / is hydrogenated in the same manner as in Example 5 step c /, and the product is isolated as the oxalate salt (Yield 90%).

 P.F .: 90-920C.

1 H-NMR (CD30D, ppm): 7.00-7.25 (4H, m, ArH); 5.30 (1H, m, NCHCO); 4.8-5.0 (2H, m, CH2 isoindoline); 3.25 (1H, m,
NCHCO); 2.90-3.00 (2H, m, CH 2 N); 1.50 - 2.00 (2H, m, CH 2 aminobutyryl); 0.8-1.2 (10H, m, CH 3 CH 2 CH 2 CH 3 aminobutyryl).

Microanalysis: C17H25N3O2.C2H2O4.1,25H2O
Found: C, 54.98%; H, 7.30%; N = 10.66%
Calculated: C = 54.86%; H, 7.15%; N = 10.10%
Example 34: Preparation of 1- (2 (S) -aminobutyryl) (R / S) -nipecotamide (UCL 1378-H); lane 3
R1 = CH2CH3; R2 = H; n = 1 and m = 0
R3 = - (CH2) 2-CH (CONH2) -; R4 = H
The procedure is as for the previous examples according to route 3.

PF = 149-150 ° C
1 H-NMR (200 MHz, CD3 OD, ppm): 4.3-4.5 (2H, m, 2 x CH) 3.6-4.0 (2H, m, CHCH2N nipecotamide); 2.8-3.1 (2H, m, CH 2 CH 2 Nipecotamide); 1,4-2,1 (6H, m, CH 2 CH 2 CH nipecotamide, CH 2 aminobutyryl); 1.00 (3H, m, CH 3 aminobutyryl).

 IR (nujol, cm-1): 1570 (N-H amide); 1650 (C = O amide).

Example 35: Preparation of L-valyl-L-proline cyclohexylmethylamide (UCL 1331-J); lane 1
R1 = CH (CH3) 2; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-NH-R5; R5 = CH2C6H11
The procedure is as for the other examples of proline backbone compounds prepared according to route 1 and the title compound is obtained in the oxalate salt form.

Mp = 157-158 ° C
IR (nujol, cm-1): 3369-3265 (NH); 3600-2400 (OH); 2935 (nujol); 1745-1635 (C = O); 1459-1375 (nujol).

1 H-NMR (200 MHz, DMSO-d6, ppm): 7.85-7.9 (1H, t, NH) 4.28-4.35 (1H, q, oeH proline); 3.94-3.97 (1H, d, aH valine) 3.5-3.7 (2H, m, -CH2N-proline); 2.82-2.95 (2H, m, -NH-CH2
C6H1l); 1.51-2.50 (10H, m, -NCH2CH2CH2-proline, CH (CH3) 2, -CH2CHCH2-cyclohexyl); 0.74-1.4 (12H, m, -CH 2 CH 2 CH 2 -cyclohexyl and 2 x CH 3 -).

Microanalysis: C17H31N3O2 (COOH) 2.O, 2H2O:
Found: C, 56.87%; H, 8.42%; N = 9.98%
Calculated C = 56.61%; H, 8.35%; N = 10.42%
Example 36: Preparation of L-valyl-L-proline benzylamide (UCL 1353-H); lane 1
R1 = CH (CH3) 2; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-NH-R5; R5 = -CH2Ph
The procedure is as for the other proline backbone compounds prepared according to route 1 and the title compound is obtained in the oxalate salt form.

PF = 143-144 ° C
IR (KBr, cm-1). 3111 (CH); 3073 (CH); 2968 (CH) 1695 (C = O); 1670 (C = O); 1639 (C = O); 1618-1556-1498 (C = C in
Ph).

 1 H-NMR (200 MHz, DMSO-d6, ppm): 8.50-8.53 (1H, m, NH) 7.30-7.34 (5H, m, aromatic H); 4.24-4.39 (3H, m, Ph-CH 2, aCH proline); 3.97-4.0 (1H, m, cCH valine); 3.45-3.99 (2H, m, -NCH2-proline) 1.76-2.17 (5H, m, -NCH2CH2CR2-proline, ssCH valine); 0.90-1.50 (6H, m, 2 x CH 3).

Microanalysis: C17H25N3O2.C2H2O4.0,2H20
Found: C, 57.42%; H, 6.94%; N = 10.48%
Calculated: C, 57.48%; H = 6.96%; N = 10.58 *
Example 37 Preparation of 1- (2 (S) -aminobutyryl)
L-Proline (R) sec.butylamide (UCL 1387-J); lane 1
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-NH-R5 R5 = -CH (CH3) CH2CH3
The procedure is as for the other proline backbone compounds prepared in lane 1 and the title compound is obtained as the oxalate salt.

Mp = 165-166oC
IR (KBr disc, cm-1): 3650-3150 (OH); 3000-2900 (CH) 1710 (C = O); 1650 (C = O); 1200 (COC).

1 H-NMR (200 MHz, DMSO-d6, ppm): 7.58-7.64 (1H, d, NH) 4.18-4.38 (1H, m, aCH); 4.0-4.12 (1H, m, &alpha;CH); 3.28-3.76 (3H, m, -NCH2-proline, -NH-CH-sec.butyl); 1.54-2.24 (6H, m, -NCH 2 CH 2 CH 2 -proline, CH 2 CH 3 aminobutyryl); 1.27-1.50 (2H, m,
CH2); 0.72-1.1 (9H, m, 3 x CH3).

Microanalysis: C13H25N3O2.0,93C2H2O4.0,6H2O
Found: C, 51.22%; H, 7.96%; N = 11.79%
Calculated: C, 51.01%; H, 8.08%; NN = 12.01%
Example 38 Preparation of 1- (2 (S) -aminobutyryl)
L-proline (S) sec.butylamide (UCL 1416-H); lane 1
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-NH-R5; R5 =
CH (CH3) CH2CH3
The procedure is as in the preceding example and the title compound is obtained in the form of an oxalate salt.

Mp = 153-154 ° C
1 H-NMR (200 MHz, DMSO-d 6, ppm) 7.61-7.65 (1H, d, NH) 4.22-4.28 (1H, m, CH) 3.97-4.03 (1H , m, &alpha;CH); 3.30-3.68 (3H, m, NCH2-proline, NH-CH-); 1.58-2.13 (6H, m, ssCH2aminobutyryl, NCH2CH2CH2-proline); 1.23-1.39 (2H, m,
CH2); 0.71-1.02 (9H, m, 3 x CH3).

Microanalysis: C13H25N3O2.0,9C2H2O4.0,2C2H6O
Found: C, 52.72%; H, 8.31%; NN = 12.28 96
Calculated: C = 52.83%; H, 8.17%; N = 12.16%
Example 39: Preparation of 2- (2 (S) -aminobutyryl) 1,2,3,4-tetrahydro-3 (S) -isoquinolinecarboxylic acid methylamide (UCL 1485-H); lane 3
R1 = CH2CH3; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 =

R4 = CO-NH-R5; = = CH3
The procedure is analogous to Example 18 and the title compound is obtained as the oxalate salt.

Mp 119-124 ° C
1 H-NMR (400 MHz, ppm): 7.29-7.19 (4H, m, ArCH); 4,564.85 (masked with H 2 O, m, NCH 2 Ar, 2 x aCH); 3.06-3.22 (2H, m,
ArCH2CH); 2.68 (3H, s, NHCH3); 1.96-2.06 (2H, m, CH 3 CH 2) 1.11-1.14 (masked, m, CH 3 CH 2).

Microanalysis: C15H21N302.1.0 (COOH) 2.0.5R20.0.75C2HsOH
Found: C, 54.48%; H, 6.80%; N = 10.05%
Calculated: C, 54.28%; H, 7.19. N = 9.99%
MEASUREMENT OF MEMBRANE TRIPEPTIDYLPEPTIDASE ACTIVITY
After isolating and characterizing the above-mentioned membrane tripeptidylpeptidase, the inventors have developed a measure of its activity described below.

 Rat brain membranes were obtained by centrifugation (200,000 x g min) of a homogenate prepared in 10 volumes of 50 mM potassium phosphate buffer, pH 7.4.

 The thoroughly washed centrifugation pellet is taken up in the buffer containing 10% glycerol and 0.1% Brij to obtain a concentration of 25 mg protein / ml.

 After 25 minutes of preincubation, the incubations are carried out for 15 minutes at 37 ° C. in the buffer in a volume of 0.1 ml containing 0.1% Brij 35, 100 mM bestatin, 25 μg of membrane and 50 μM of substrate (A-AP-Amc).

The release of Amc is evaluated by fluorometry.

DEVELOPMENT OF INHIBITORS
The effect of the compounds according to the invention given as examples above was evaluated by measuring the membrane tripeptidylpeptidase II activity. The inhibitory power of these compounds was expressed by their apparent dissociation constant (Ki), calculated from the values of their 50% inhibitory concentration and the KM of the substrate (23 μM).

 The following Table II reports a series of characteristic values of the compounds according to the invention.

TABLE II
INHIBITION OF AMINOTRIPEPTIDYLPEPTIDASE ACTIVITY
BRAIN MEMBRANE
Internal code of compound n "example Ki vM
UCL 1330-J 1 0.9
UCL 1382-J 2 0.3
UCL 1391-J 3 0.270
UCL 1371-H 4 0.10
UCL 1381-J 5 0.5
UCL 1393-F 6 0.080
UCL 1394-F 7 0.320
UCL 1327-J 8 57
UCL 1324-J 9 0.57
UCL 1372-F3 10 0.00030
UCL 1389-J 11 0.00030
UCL 1419-H 12 5.0
UCL 1420-H 13 12.0
UCL 1424-A 14 5.0
UCL 1395-F 15 0.320
UCL 1433-H 16 0.140
UCL 1374-E 17 40
UCL 1421-H 18 0.040
UCL 1482-H 19 6.0
UCL 1398-F 20 12.0
UCL 1373-E 21 20
UCL 1397-H 22 0.0052
UCL 1413-H 23 0.0060
UCL 1414-H 24 0.340
UCL 1415-H 25 0.012
UCL 1423-H 26 8.0
UCL 1446-A2 27 2.8
UCL 1486-H 28 0.008
UCL 1546-H 29 0.077
UCL 1534-H 30 0.012
UCL 1535-A 31 1.0
UCL 1442-H 32 0.560
UCL 1466-H 33 0.143
UCL 1378-H 34 40
UCL 1331-J 35 34
UCL 1353-H 36 3.3
UCL 1387-J 37 4,5
UCL 1416-H 38 20
UCL 1485-H 39 12
UCL 1338-J * known 0.57
UCL 1702-H 30 0.003
UCL 1703-H 30 0.233
UCL 1704-H 28 0.006
UCL 1705-H 28 0.241 * 2 (S) -aminobutyryl-L-prolinamide
R1 = CH (CH3) 2; R2 = H; n = 0 or 1 and m = 0 or 1 with n different from m; R3 = - (CH2) 2-; R4 = CO-NH-R5; R5 = H
Remarkable effects of the compounds according to the invention have been demonstrated in particular for compounds having an indoline backbone. It is notable that the compound referenced under the UCL 1397-H internal code, for example, has an endogenous CCK-8 protective effect on depolarized rat cerebral cortex section as evidenced by the method of Rose et al.

(Proc Natl Acad Sci USA, 1988, 85: 8326). At a concentration of 1 μM, it multiplies by 4 the recovery of
CCK-8 released (measured by radioimmunoassay).

 The anorexigenic activity (pro-satiety) of this same compound is demonstrated in rodents: in the mouse, at a dose of 10 mg / kg (intravenously), this compound reduces by 45% the spontaneous intake of food solid (tablet "UAR") measured over a period of 30 to 90 minutes after free access to this food (period preceded by overnight fasting and administration of 0.2 ml of a semi-liquid meal by mouth ).

 This test is completed by an evaluation of the "ex vivo" TPP II inhibitory activity of administering a compound according to the invention to a rodent and measuring residual catalytic activity on brain membranes or peripheral organs.

For example, hepatic TPP II activity measured in the mouse 90 minutes after administration of 10 mg / kg of the compound referenced under the UCL 1372-F3 internal code orally is inhibited to about 90-100%.

SEQ ID1 cgcccctcgtccgcgcgctgcctggcagtttacctcttccacgtccgtcctccagcttgc 60 gtcc ATG GCC ACC GCT GCG ACC GAG GAG CCT TTC CCT TTC CAT GGT 106
MATAATEEPFPFHG 14
CTT CTA CCA AAG AAA GAG ACT GGG GCC TCC TCC TTC CTG TGC CGC 151
LLPKKETGASSFLCR 29
TAC CCG GAG TAT GAC GGA CGC GGG GTG CTC ATC GCC GTC CTG GAC 196
YPEYDGRGVLIAVLD 44
ACA GGG GTT GAT CCC GGG GCC CCG GGC ATG CAG GTC ACG ACT GAT 241
TGVDPGAPGMQVTTD 59
GGG AAA CCA AAA ATT ATC GAT ATC ATT GAT ACA ACA GGA AGT GGT 286
GKPKIIDIIDTTGSG 74
GAT GTG AAT ACT GCT GAA GAA GTA GAA GAA AAA GAT GGC GAA ATC 331
DVNTATEVEPKDGEI 89
ACT GGT CTT TCT GGA AGA GTG CTT AAG ATT CCT GCA AAC TGG ACA 376
TGLSGRVLRIPANWT 104
AAT CCT TCA GGA AAA CAT TAT ATT GGC ATT AAA AAT GGT TAT GAC 421 NPSGKYHIGIKNGYO 119
TTC TAT CCA AAG GCT CTC AAG GAA AGG ATA CAG AAA GAA CGG AAG 466 FYPKALKERIQKERK 134
GAA AAA ATC TGG GAT CCA ATT AGC AGA GTT GCG CTT GCA GAA GCT 511
EKIWDPIHRVALAEA 149
TGT AGA AAA CAA GAA GAA TT GAT ATT GCC AAT AAT GGC TCT CBT 556
CRKQEEFDIANNGSS 164
CAA GCC AAT AAA CTA ATC AG GAA GAG TTG AG AG AGA CAA GTG GAA 601
QANKLIKEELQSQVE 179
TTA CTT AAT TCT TT GAG AAA AGT TAT AGT GAT CCA GGC CCT GTG 646
LLNSFEKKYSD :: GPV 194
TAT GAC TGC TTG TGG TGG GAT CAT GGT ACC GAG TG AGA GCC TGT 691
YDCLVWHDGETWRAC 209
GTT GAT TCG AAT GAA AAT GGG GAC TTG GGT AAA TCT ACT GTA TTG 736 VOSNENGO t GKSTV t 224
AGA AAC TAC AAA GAG GCT CAA GAAT TAT GGT TCT TTT GGC ACA GCT 781
RNYKEAQEYGSFGTA 239
GAG ATG CTG AAT TAC TCT GTT AAT ATT TAT GAT GAT GGA AAC CTG 826 EM t NYSVNIYODGN t 254
CTC TCC ATT GTG ACC AGT GGA GGA GCT GGA CAT ACC CAT GTA GCG 871 LSIVTSGGAHGTHVA 269
AGT ATA GCT GCT GGG CAT TT CCA GAA GAG CCT GAA CGG AAT GGA 916
SIAAGHFPEEPERNG 284
GTT GCT CCT GGT GCT CAA ATT CTA TCC ATT GG ATTAT ATT GAT ACA 961
VAPGAQILSIKIGDT 299
AGG CTA AGC ACC ATG GAA ACA GGC ACA GGC CTC ATC AGA GCT ATG 1006
RLSTMETGTGLIRAM 314
ATA GAA GTT ATA AAT CAT AAG TGT GAT CTT GTC AAC TAC AGT TAT 1051 IEVINHKCD t VNYSY 329
GGA GAA GCA CAT ACT TGG CCA AAT TCT GGG ATT AGT TGT GAA GTA 1096 GEATHWPNSGRICEV 344
ATT AAT GAA GCA GTA TGG AAA AAT ACA ATT TAT GTT TCA AGT 1141
INEAVWKHNTIYVSS 359
GCT GGA AAT AAT GGT CCA TGC CTT TCT ACA GTG GGT TGT CCA GGA 1186 AGNNGPC t STVGCPG 374
GGA ACT ACT TCC AGC GTG ATA GGT GGA GTT GTT TAT GTT TCT CCT 1231 GTTSSVIGVGAYVSP 389
GAT ATG ATG GTT GCA GAG TAT TCA CTG AGA GAG AAA CTA CCT GCA 1276 DMMVAEYS t REKLPA 404
AAT CAA TAT ACA TGG TCT TCT AGA GGC CCA AGT GCT GAT GGG GCC 1321 NQYTWSSRGPSADGA 419
CTT GGT GTG AGC ATC AGT GCA CCA GGA GGT GCT ATT GCT TCT GTG 1366 t GVSISAPGGAIASV 434
CCT AAC TGG ACA TTG AGG GGG ACT CAG CTA ATG AAT GGG ACA TCA 1411 PNWT t RGTQ t MNGTS 449
ATG TCT TCC CCC AAT GCC TGT GGT GGC ATT GCC CTG GTA CTT TCA 1456 MSSPNACGGIA t VLS 464
GGG CTG AAA GCA AAT AAT GTT GAC TAT ACT GTT CAC TCA GTC AGA 1501 G t KANNVDYTVHSVR 479
AGA GCT CTA GAA AAC ACT GCA ATA AAA GCT GAC AAT ATA GAA GTA 1546 RA T ENTAIKADNIEV 494
TTT GCC CAA GGA CAT GGA AT ATT CAG GTT GAC AAA GCT TAT GAC 1591 FAQGHGIIQVDKAYD 509
TAC CTC ATT CAG AAT ACA TCA TT GCT AAC AGA TTA GGT TT ACA 1636
YT IQNTSFANR t GFT 524
GTT ACA GTT GGA AAT AAC CGT GGC ATC TAC CTC CGA GAT CCT GTT 1681 VTVGNNRGIY t RDPV 539
CAG GTG GCT GCT CCT TCA GAT CAT GGT GTT GGC ATT GAG CCT GTA 1726 QVAAPSDHGVGIEPV 554
TTT CCA GAG AAC ACA GAA AAC TCT GAA AAA ATA TCC TTC CAG CTT 1771 FPENTENSEKISFQ t 569
CAT TTA GCT TTA ACT TCA AAT TCA TCT TGG GTT CAA TGT CCC AGC 1816
HLALTSNSSWVQCPS 584
CAT TTG GAA CTC ATG AAT CAA TGT CGG CAC ATA AAC ATA CGT GTG 1861 H T E T MNQCRHINIRV 599
GAC CCC AGG GGC TTA AG GAA GGG TTA CAT TAT AC GAG GTA TGT 1906
DPRGLREGLHYTEVC 614
GGG TAT GAT ATA GCA TCC CCC AAT GCA GGT CCT TTA TTC AGA GTT 1951 GYDIASPNAGP t FRV 629
CCA ATC ACT GCA GTT ATA GCA GCA AAA GTA AAT GAA TCA TCA CAT 1996 PITAVIAAKVNESSH 644
TAT GAT CTA GCC TT ACA GAT GTA CAT TT AAA CCT GGT @AG A @@ 2041
YDLAFTDVHFKPGQI 659
CGA AGA CAT TTT GTT GAG GTT CCT GAG GGG ACT GCA TGG GCT GAA 2086
RRHFVEVPEGATWAE 674
GTT ACC GTG TGT TCG TGT TCT TCT GAG GTA TCA GCA AAA TTT GTT 2131
VTVCSCSSEVSAKFV 689
CTT CAT GCA GTA AGC CTT GTG AGC AG AGA GCA AGC TAT AGT CAT 2176
LHAVQLVKQRAYRSH 704
GAA TTT TAT AAG TT TGT TCC CTT CCA GAA AAA GGA ACA CTC ATT 2221
EFYKFCSLPEKGTLI 719
GAA GCC TT CCT GTT CTG GGT GGG AAA GAA ATT GAA TT TGT ATT 2266 EAFPVLGGKAIEFCI 734
GCT CGT TGG TGG GCA AGT CTC AGT GAT GTC AAT ATT GAT TAT ACC 2311
AKWWASLSDVNIDYT 749
ATA TCA TTC CAT GGC ATA TGG TGT ACT GCA CCA AGC TTA AAC ATT 2356
ISFHGIVCTAPQLNI 764
CAT GCA TCT GAA GGA ATC AAT CGT TT GAT GTC AGC TCC TCT TTA 2401
HASEGINKFDVQSSL 779
AAA TAT GAA GAT CTG CCT GCT TGC ACT ATA TTG AGC AGC TGG GTG 2446
KYEDLAPCITLKSWV 794
CAA ACA CTA CGC CCA GTA AAA GCA AAA ACC AGA CCT TTA GGA TCA 2491
QTLRPVNAKTRPLGS 809
AGA GAT GTT TTG CCA AAT AAT CGC CAG CTT TAT GAG ATG GTC CTG 2536
RDVLPNNRQLYEMVL 824
ACA TAC AGC TT CAT CAG CCC AAG AGC GGA GAA GTA ACA CCT AGT 2581
TYSFHQPKSGEVTPS 839
TGT CCA CTC CTT TGT GAA TTA TTA TAT GAG TCA GAA TTT GAC AGT 2626
CPLLCELLYESEFDS 854
CAG TTG TGG ATT ATT TTT GAC CAG AAC AAA AGA CAG ATG GGC TCA 2671
QLWIIFDQNKRQMGS 869
GGC GAT GCC TAT CCA CAT CAG TAT TCT TTG AAA TTG GAG AAA GGA 2716
GDAYPHQYSLKLEKG 884
GAT TAT ACG ATT CGA TTA CAG ATT CGT CAT GAG CAA ATC AGT GAT 2761 DYTIRLQIKHEQISD 899
TTG GAT CGT CTC AAA GAT CTT CCG TT ATT GTT TCG CAT AGG TTG 2806
LDKLKD t PFIVSHKL 914
TCT AAT ACC TTG AGC TTA GAT ATT CAT GAA AAT AGC CAT CTT GCA 2851 SNT t S t DIHENHS t A 929
CTT CTA GGA AAG AAA AAA TCA AGC AGT TTG ACA TTA CCA CCC AAA 2896
tt GKKKSSS t T t PPK 944
TAT AAT CAG CCA TTC TT GTT ACT TCC TTA CCT GAT GAT AAA ATA 2941 YNQPFFVTS t PDDKI 959
CCT AAG GGG GCA GGA CCC GGA TGC TAC CTT GCA GGC TCC TTG ACA 2986 PKGAGPGCY t AGS t T 974
TTG TCA AAG ACT GAG CTT GGA AAA AAA GCT GAT ATC GTG (2733995 3031 LSKTELGKKADVIPV 989
CAT TAC TAT CTA ATA CCT CCA CCA ACA AGA ACT AAG AGG AGC 3076 HYYLIPPPTKTKNGS 1004
AAA GAAT AAA GAA AAA GAT GAA GAA AAA AAA GAG GAAT TTG AAA GAA 3121 KDKEKDSEKEKDLKE 1019
GAG TTT ACT GAA GCT TTA CGA GAT CTT AAA ATT CAG TGG ATG ACC 3166
EFTEALRDLKIQWMT 1034
AAG CTG GAT TCT ACT GAC AT ATT ATTACLE AAT GAA TTG GAA GAA ACA TAT 3211 KLDSTDIYNELKETY 1049
CCT GCT TAC CTT CCT TTG TAT GTT GCA CGT CTT CAT CAA TTA GAT 3256 PAYLPLYVARLHQLD 1064
GCT GAA AAG GAG CGG ATG AAA AGM CTT AAT GAA ATT GTT GAT GCT 3301 AEKERMKRLNEIVDA 1079
GCC AAT GCT GTT ATT TCT CAC ATC GAT CAG ACC GCT CTC GCT GTT 3346 ANAVISHIDQTALAV 1094
TAC ATT GCC ATG AAG ACT GAC CCC AGG TCC GAT GCA GCT ACT ATA 3391 YIAMKTDPRPDAATI 1109
AAA AAT GAT ATG GAC AAG CAG AAA TCT ACC CTG GTA GAT GCC CTC 3436 KNDMDKQKSTLVDAL 1124
TGC AGG AAA GGA TGT GCT CTG GCA GAC CAC CTT CTT CAT GCA CAG 3481 CRKGCALADHLLHAQ 1139
CCC CAC GAT GGG GCA GCA GCT GGA GAT GCA GAA GCA AAA GAA GAG 3526
PHDGAAAGDAEAKEE 1154
GAA GGA GAA AGT ACC TTG GAA TCT CTA TCA GAA ACC TAT TGG GAA 3571 EGESTLESLSETYWE 1169
ACA ACT ACG TGG ACA GAT CTT TT GAC ACT AAG TTG TTG ACA TT 3616 TTKWTO t FDTKV t TF 1184
GCA TAC AAG CAT GCA TTA GTA AAT AAG ATG TAC GGG AGA GGC CTT 3661 AYKHA t VNKMYGRG t 1199
AAG TT GCA ACC AAA CTC GTA GAA GAA AAA CCA ACA AAA GAA AAC 3706 KFATK t VEEKPTKEN 1214
TGG AAA AAT TGT ATT CAA CTG ATG AAA TTA CTC GGA TGG ACC CAT 3751
WKNCIQLMKLLGWTH 1229
TGT GCG TCT ACT TTT GAA AAC TGG TCC CCC ATC ATG TAT TAC CCT 3796
CASFTENW t PIMYPP 1244
GAT TAT TGT GTA TAA TTC aatgggaaccaaaacgttaaatttcgaaagcagaa 3849
DYCVF aattttatagtgaacaaatatatgaacaaatgtgtggcatttctagtctaactaatgcat * 1250 3909 3969 gtcttcatccactatcgaatactgatcattaaaactctatgtatttatcagagaactcaa tggtgtgtggcttcatacatgtaatgtagacagacctctgacatcatgctgctttcctgc tgcctcccacacttggctagggagggcagagcctgcctgccagccccaacctggtggat 4029 4089 4149 gcagctgctcactgcaggagaggttttcatctcttaattttaactgtaaaacgtcatcc agtttttattttataaatcaaaaaggttaaaacatgctaaatttttccaattat tagag gccttaaaaatgctacattgggtgtagctaaattatttatttgactaaaaattacgagaa 4209 4269 4329 cataatttccagacttctaaaaataaattcaattaatgggtatggtggggaggtataaat acatggcaactgggaaaagaaacgtgttaatgtaatcttcactccggagtcacaacaagc aagttgtttttacagcatcctcaagtacacagcatcagaataagttaaaatttcatgtgt 4389 4449 4509 tggtgccagacagttgaatctatctggttttgtaaagatatacacagtatgtttataaca ttgaaatcatgtaaaatacatgaataaatgtgcaaaacaacaggcacagcacaccatatg cactctgatacctgttttttataaataaaagtaaatattgaagctaaa 4569 4617 SEQ @D @ ataatttgtttggtttcactgctgagtaccatgctatgatttctacactatctgcttatt 60 aaactgtttcttactgtttggaactgttatctaacagttatttcaaaaagcattaggcac 120 agttc ctgtgcctgtgtgcagatgtttccaggatgtataccagaaatggaattgctgagt cagggagattcaggtttactaggtcatgtgtaacagctttttcattgagctggccttgaa 180 240 300 ctcttctgatattcctgcctccacctcccagctgctgtgattatggatatattccaccat gcctggccaactgttaacttcttttttttaattttattttttttttaaagattttatttt 360 attgtatgagtgcactgtagctgccttcagacacaccagaagagggcattggattccatt 420 acagatggttgtgagccaccatgtggttgctgggatttgaactcaggacctcaggaagag cactcggtgctcttaaccactgagccatctctccagcccccaactgttaacttcttatac 480 540 598 tttgaattttcagacctgtttttaatttcttatgtaggaaatatcagtgtattgtagg
TTT GAT TTG CAT TTT TAT AGT GAA ATC AGT TAT TTT TCT AGA AAC 643
FOLHFYSEISYFSNR 15
TTT AGG CTT TTG GGT CCT CTC ATA GTG CTA TTA AAG TTC TTT TAC 688
FRLLGPLIVLLKFFY 30
TTT TTC ATT TTC TGT TT TT TT TT TT TT ACT ACT ACT TTG 733
FFIFCFFFFFNTTKL 45
TCA ATT GCT TTT TTT TTT TTT CCG GAG CTG GGG ATC GAA CCC AGG 778
SIAFFFFPE t GIEPR 60
GCC TTG CGC TTG CTA GGC AAG CGC TCT ACC GCT GAG CTA AAT CTC 823
ALRLLGKRSTAELNL 75
CAA CCC CTG ATT TC GCT TT TCA CTG ATT CCT GCA TCA TGG ACA
QPLSIAFSLIPANWT
AAT CCT TCA GGA AAA CAT TAT ATT GGC ATT AAA AAT GGT TAT GAC NPSGKYH t GIKNGYD
TTC TAT CCA AAG GCT CTC AAG GAA AGG ATA CAG AAA GAA CGG AAG
FYPKALKERIQKERK
GAA AAA ATC TGG GAT CCA ATT AGC AGA GTT GCG CTT GCA GAA GCT
EKIWDPIHRVALAEA
TGT AGA AAA CAA GAA GAA TTT GAT ATT GCC AAT AAT GGC TCT TCC
CRKQEEFDIANNGSS
CAA GCC AAT AAA CTA ATC GAA GAG TTG CAG AGT CAA GTG GAA
QANKLIKEELQSQVE
TTA CTT AAT TCT TT GAG AAA AGT ATAT AGT GAT CCA GGC CCT GTG
LLNSFEKKYSDPGPV
TAT GAC TGC TTG TGG TGG GAT CAT GGT ACC GAG TGG AGA GCC TGT
YDCLVWHDGETWRAC
GTT GAT TCG AAT GAA AAT GGG GAC TTG GGT AAA ACT TCT GTA TTG VDSNENGDLGKSTVL
AGA AAC TAC AAA GAG GCT CAA GAA TAT GGT TCT TTT CCC ACA GCT
RNYKEAQEYGSFGTA
GAG ATG CTG AAT TAC TCT GTT ATTAT TAT GAT GAT GGA AAC CTG
EMLNYSVNIYDDGNL
CTC TCC ATT GTG ACC AGT GGA GGA GCT GGA CAT ACC CAT GAT CCC
LSIVTSGGAHGTHVA
AGT ATA GCT GCT GCC CAT TTT CCA GAA GAG CCT GAA CCC AAT GGA
SIAAGHFPEEPERNG
GTT GCT CCT GGT GCT CAA ATT CTA TCC ATT AAC ATT GGT GAT ACA
VAPGAQILSIKIGDT
AGG CTA AGC ACC ATG GAA ACA GGC ACA GGC CTC ATC AGA GCT ATG RLSTMETGTGLIRAM
ATA GAA GTT ATA AAT CAT AAG TGT GAT CTT GTC AAC TAC AGT @@@
IEVINHKCDLVNYSY
GGA GAA ACT GCA CAT TGG CCA AAT TCT GGG ATT AGT TGT GAA GTA
GEATHWPNSGRICEV
ATT AAT GAA GCA GTA TGG AAA AAT ACA ATT TAT GTT TCA AGT
INEAVWKHNTIYVSS
GCT GGA AAT AAT GGT CCA TGC CTT TCT ACA GTG GGT TGT CCA GGA AGNNGPO t STVGOPG
GGA ACT ACT TCC AGC GTG ATA GGT GGA GTT GCT TAT GTT TCT CCT
GTTSSVIGVGAYVSP
GAT ATG ATG GTT GCA GAG TAT TCA CTG AGA GAG AAA CTA CCT GCA DMMVAEYS t REK t PA
AAT CAA TAT ACA TGG TCT TCT AGA GGC CCA AGT GCT GAT GGG GCC
NQYTWSSRGPSADGA
CTT GGT GTG AGC ATC AGT GCA CCA GGA GGT GCT ATT GCT TCT GTG t GVSISAPGGAIASV
CCT AAC TGG ACG TTG AGG GGG ACT CAG CTA ATG AAT GGG ACA TCA
PNWTLRGTQLMNGTS
ATG TCT TCC CCC AAT GCC TGT GGT GGC ATT GCC CTG GTA CTT TCA MSSPNACGGIA t V t S
GGG CTG AAA GCA AAT AAT GTT GAC ACT TAT GTT CAC TCA GTC AGA
GLKANNVDYTVHSVR
AGA GCT CTA GAA AAC ACT GCA ATA AAA GCT GAC AAT ATA GAA GTA RA T ENTAIKAONIEV
TTT GCC CAA GGA CAT GGA AT ATT CAG GTT GAC AAA GCT TAT GAC FAQGHGIIQVDKAYD
TAC CTC ATT CAG AAT ACA TCA TT GCT AAC AGA TTA GGT TT ACA
YLIQNTSFANRLGFT
GTT ACA GTT GGA AAT AAC CGT GGC ATC TAC CTC CGA GAT CCT GTT VTVGNNRGIY t ROPV
AGC GTG GCT GCT CCT TCA CAT GGT GTT GGC ATT GAG CCT GTA
QVAAPSDHGVGIEPV
TTT CCA GAG AAC ACA GAA AAC TCT GAA AAA ATA TCC TTC CAG TTC
FPENTENSEKISFQL
CAT TTA GCT TTA ACT TCA AAT TCA TCT TGG GTT CAA TGT CCC AGC
HLALTSNSSWVQCPS
CAT TTG GAA CTC ATG AAT CAA TGT CGG CAC ATA AAC CGT CGG
HLELMNQCRHINIRV
GAC CCC AGG GGC TTA AGA GAA GGG TTA CAT TAT ACA GAG GTA TGT OPRG t REG t HYTEVC
GGG TAT GAT ATA GCA TCC CCC AAT GCA GGT CCT TTA TTC AGA GTT GYOIASPNAGP t LIF
CCA ATC ACT GCA GTT ATA GCA GCA AAA GTA AAT GAA TCA TCA CAT p ITAVIAAKVNESSH
TAT GAT CTA GCC TT ACA GAT GTA CAT TT AAA CCT GGT (2733995
YDLAFTDVHFKPGQI
CGA AGA CAT TTT GTT GAG GTT CCT GAG GGG ACT GCA TGG GCT GAA
RRHFVEVPEGATWAE
GTT ACC GTG TGT TCG TGT TCT TCT GAG GTA TCA GCA AAA TTT GTT
VTVCSCSSEVSAKFV
CTT CAT GCA GTA CAG CTT GTG CAG AGA GCA TAT CGA AGT CAT
LHAVQLVKQRAYRSH
GAA TTT TAT AAG TT TGT TCC CTT CCA GAA AAA GGA ACA CTC ATT
EFYKFCSLPEKGTLI
GAA GCC TT CCT GTT CTG GGT GGG AAA GCA ATT GAA TT TGT ATT EAFP vt GGKAIEFCI
GCT CGT TGG TGG GCA AGT CTC AGT GAT GTC AAT ATT GAT TAT ACC ARWWAS SDVNIDYT
ATA TCA TTC CAT GGC ATA GTG ACT TGC CCA AGC TTA AAC ATT
ISFHGIVCTAPQLNI
CAT GCA TCT GAA GGA ATC AAT CGT TTT GAT GTC CAG TCC TCT TTA HASEGINRFDVQSSL
AAA TAT GAA GAT CTG CCT GCT TGC ACT ATA TTG AGC AGG TGG GTG KYEO t APCIT t KSWV
CAA ACA CTA CGC CCA GTA AAT GCA AAA ACC AGA CCT TTA GGA TCA
QTLRPVNAKTRPLGS
AGA GAT GTT TTG CCA AAT AAT CGC CAG CTT TAT GAG ATG GTC CTG RDVLPNNRQLYEMVL
ACA TAC AGC TT CAT CAG CCC AAG AGC GGA GAA GTA ACA CCT AGT
TYSFHQPKSGEVTPS
TGT CCA CTC CTT TGT GAA TTA TTA TAT GAG TCA GAA TTT GAC AGT CPLLCELLYESEFDS
CAG TTG TGG ATT ATT TTT GAC CAG AAC AAA AGA CAG ATG GGC TCA
QLWIIFDQNKRQMGS
GGC GAT GCC TAT CCA CAT CAG TAT TCT TTG AAA TTG GAG AAA GGA
GDAYPHQYSLKLEKG
GAT TAT ACG ATT CGA TTA ATT ATT CGT CAT GAG CAA ATC AGT GAT
DYTIRLQIRHEQISD
TTG GAT CGT CTC AAA GAT CTT CCG TT ATT GTT TCG CAT AGG TTG LDRLKDLPFIVSHRL
TCT AAT ACC TTG AGC TTA GAT CAT GAA ATTAT CAT AGC CTT GCA
SNTLSLDIHENHSLA
CTT CTA GGA AAG AAA AAA TCA AGC AGT TTG ACA TTA CCA CCC AAA
LLGKKKSSSLTLPPK
TAT AAT CAG CCA TTC TT GTT ACT TCC TTA CCT GAT AAA GTA ATA
YNQPFFVTSLPDDKI
CCT AAG GGG GCA GGA CCC GGA TGC TAC CTT GCA GGC TCC TTG ACA
PKGAGPGCYLAGSLT
TTG TCA AAG ACT GAG CTT GGA AAA AAA GCT GAT GTG ATC
LSKTELGKKADVIPV
CAT TAC TAT CTA ATA CCT CCA CCA ACA AAG ACT AAG AAT GGC AGC
HYYLIPPPTKTKNGS
AAA GAAT AAA GAA AAA GAT GAA GAA AAA AAA GAG AAA AAA GAA GAA GAKE KOKEKDSEKEKDLKE
GAG TT ACT GAA GCT TTA CGA GAT CTT AAA ATT AGC TGG ATG ACC
EFTEALRDLKIQWMT
AAG CTG GAT TCT ACT GAC ATT TAC AAT GAA TTG AAA GAA ACA TAT
KLDSTDIYNELKETY
CCT GCT TAC CTT CCT TTG TAT GTT CGC CGT CTT CAT CAA TTA GAT
PAYLPLYVARLHQLD
GCT GAA AAG GAG CGG ATG AAA AGM CTT AAT GAA ATT GTT GAT GCT AEKERMKR t NEIVOA
GCC AAT GCT GTT ATT TCT CAC ATC GAT CAG ACC GCT CTC GCT GTT ANAVISHIDQTALAV
TAC ATT GCC ATG AAG ACT GAC CCC AGG TCC GAT GCA GCT ACT ATA
YIAMKTDPRPDAATI
AAA AAT GAT ATG GAC AAG CAG AAA TCT ACC CTG GTA GAT GCC CTC KNDMDKQKSTLVDAL
TGC AGG AAA GGA TGT GCT CTG GCA GAC CAC CTT CTT CAT GCA CAG CRKGCALADHLLHAQ
CCC CAC GAT GGG GCA GCA GCT GGA GAT GCA GAA GCA AAA GAA GAG
PHDGAAAGDAEAKEE
GAA GGA GAA AGT ACC TTG GAA TCT CTA TCA GAA ACC TAT TGG GAA
EGESTLESLSETYWE
ACT ACA AAG TGG ACA GAT CTT TTT GAC ACT AAG GTT TTG ACA TTT TTKWTO t FOTKV TF
GCA TAC AAG CAT GCA TTA GTA AAT ATG TAC GGG AGA GGC CTT AYKHALVNKMYGRGL
AAG TT GCA ACC AAA CTC GTA GAA GAA AAA CCA ACA AAA GAA AAC
KFATKLVEEKPTKEN
TGG AAA AAT TGT ATT CAA CTG ATG AAA TTA CTC GGA TGG ACC CAT WKNCIQLMKLLGWTH
TGT GCG TCT ACT ACT GAA AAC TGG CTC CCC ATC ATG TAT CCT CCT
CASFTENWLPIMYPP
GAT TAT TGT GTA TTC TAA aatgggaaccaaaacgttaaatttcgaaagcagaa OYCVF * aattttatagtgaacaaatatatgaacaaatgtgtggcatttctagtctaactaatgcat gtcttcatccactatcgaatactgatcattaaaactctatgtatttatcagagaactcaa tggtgtgtggcttcatacatgtaatgtagacagacctctgacatcatgctgctttcctgc tgcctcccacacttggctagggagggcagagcctgcctgccagccccaacctgggtggat gcagctgctcactgcaggagaggttttcatctcttaatttttaactgtaaaacgtcatcc agtttttattttataaatcaaaaaggttaaacatgctaaatttttccaattat gccttaaaaatgctacattgggtgtagctaaattatttatttgactaaaaattacgagaa cataatttccagacttctaaaaataaattcaattaatgggtatggtgggaggtataaat acatggcaactgggaaaagaaacgtgttaatgtaatcttcactccggagtcacaacaagc aagttgtttttacagcatcctcaagtacacagcatcagaataagttaaaatttcatgtgt tggtgccagacagttgaatctatctggttttgtaaagatatacacagtatgtttataaca ttgaaatcatgtaaaatacatgaataaatgtgcaaaacaacaggcacagcacaccatatg cactctgatacctgttttttataaataaaagtaaatattgaagctaaa

Claims (59)

 1 / A process for preparing a pure membrane tripeptidylpeptidase, characterized in that it comprises the following steps:  i) preparation of brain membranes, for example of rat;  ii) purification by high performance liquid chromatography (HPLC)  iii) verification of the product obtained by enzymatic reaction using a CCK substrate, for example the peptides CCK-8 (unsulfated) or CCK-5.  2 / Nucleotide sequence of a purified membrane tripeptidylpeptidase obtainable according to the method of claim 1, corresponding to the sequence identifier SEQ ID No. 1 of the attached sequence listing.  3 / nucleotide sequence of a purified membrane tripeptidylpeptidase obtainable according to the method of claim 1, corresponding to the sequence identifier SEQ ID No. 2 of the attached sequence listing.  4 / A method for screening drugs intended to treat in humans or animals disorders or conditions related to the inactivation of endogenous neuropeptides, characterized in that it consists in bringing a candidate molecule into contact with a membrane tripeptidylpeptidase or counterpart and measure the activity of this enzyme.  5 / A method according to claim 4, characterized in that the tripeptidylpeptidase is prepared according to the method of claim 1.  6 / A method according to claim 4, characterized in that the disorders or conditions are related to the inactivation of cholecystokinin (CCK). 7 / Compound of general formula (I) below

 where R6 is H, F, OCH3 or OCH2Ph,

 - R3 represents a divalent radical consisting of an alkyl chain - (CH2) p - where p represents an integer ranging from 1 to 4 (inclusive ends), -CH2-CH (cis.F) -, -CH2S -, - CH2 -CH (CH2Ph) -, - (CH2) 2-CH (CONH2) -, a pattern  - n = 0 or 1 and m = 0 or 1 with n different from m  R2 represents a hydrogen or a C1-C2 alkyl group;  R1 represents a hydrogen or a linear or branched C1-C4 alkyl group  in which

 amide CO-NH-R5 wherein R5 represents hydrogen or a linear or branched C1-C6 alkyl group, - (CH2) 3-SCH3, -CH2Ph, -CH2C6H11, CH2-NHn.C4H9,  - R4 represents a hydrogen, a group n.C6H13, CH2OH,  with the proviso that when R2 is hydrogen, R3 is the divalent radical - (CH2) 2- and R4 is the amide group CO-NH-R5, where R5 is hydrogen, then R1 is other than CH2CH3, (CH2) 2CH3 (CH2 ) 3 CH 3 CH (CH 3) 2, CH 2 CH (CH 3) 2, CH (CH 3) CH 2 CH 3. NC5H10;  or carbonyl CO-R7 where R7 is n.C5H11 or  8 / A compound according to claim 7, characterized in that R3 represents - (CH2) 2-.  9 / A compound according to claim 7, characterized in that R3 represents -CH2-CH (cis.F) -.  10 / A compound according to claim 7, characterized in that R3 represents -CH2-CH (CH2Ph) -.  11 / A compound according to claim 7, characterized in that R3 represents -SCH2-.  12 / A compound according to claim 7, characterized in that R3 represents

 13 / A compound according to claim 7, characterized in that R3 represents - (CH2) 4-.  14 / A compound according to claim 7, characterized in that R3 represents - (CH2) 3-.  15 /. Compound according to Claim 7, characterized in that R3 represents -CH2-.  16 / A compound according to claim 7, characterized in that n = 0 and m = 1.  17 / A compound according to claim 16, characterized in that R3 represents

  18 / A compound according to claim 16, characterized in that R3 represents

 19 / A compound according to claim 7, characterized in that n = 1 and m = 0.  20 / A compound according to claim 19, characterized in that R3 represents

 21 / A compound according to claim 19, characterized in that R3 represents - (CH2) 2-CH (CONH2) -.  22 / A compound according to any one of claims 7 to 20, characterized in that R4 represents CONHR5.  23 / A compound according to claim 22, characterized in that Rs represents the n-butyl group.  24 / A compound according to claim 21, characterized in that R4 represents a hydrogen.  25 / Copmposé according to claim 8, characterized in that R4 represents COR7.  26 / A compound according to claim 17, characterized in that R4 represents CH2OH.  27. Compound according to claim 17, characterized in that R4 represents CH2-NH (n.C4H9).  28 / A compound according to any one of claims 7 to 21, characterized in that R2 represents a hydrogen.  29 / A compound according to claim 8, characterized in that R1 and R2 each represent a methyl group.  30 / A compound according to claim 17, characterized in that R1 represents a hydrogen and R2 represents an ethyl group.  31 / A compound according to any one of claims 7 to 30, characterized in that it is chosen from the group comprising  2 (S) -aminobutyryl-L-prolinamide (UCL 1338-J)  - L-valyl-I, -proline n.hexylamide (UCL 1330-J)  1- (2 (S) -aminobutyryl) -L-proline 3- (methylthio) propylamide (UCL 1382-J)  1- (2 (S) -aminobutyryl) -L-proline n.pentylamide (UCL 1391 J)  1- (2 (S) -aminobutyryl) -L-proline n.butyiamide (UCL 1371 H)  1- (2 (S) -aminobutyryl) -L-proline [2 (S) -methylbutylamide (UCL 1381-J)  1- (2 (S) -aminobutyryl) -L-proline n -propylamide (UCL 1393 F)  1- (2 (S) -aminobutyryl) -L-proline iso-butylamide (UCL 1394-F)  L-valyl-L-proline n-butylamide (UCL 1324-J)  L-alanyl-L-prolyl-L-borovaline pinanediol ester (UCL 1372-F3)  L-Alanyl-L-prolyl-difluoro-L-borovaline borohydrate (UCL 1389-J)  1- (2 (S) -aminobutyryl) - (4 (S) -fluoro) -L-proline n-butylamide (UCL 1395-F)  1- (2 (S) -aminobutyryl) - (4 (S) -benzyl) -L-proline n-butylamide (UCL 1433-H)  2- (2 (S) -aminobutyryl) -1,2,3,4-tetrahydro-3 (S) -isoquinolinecarboxylic acid n-butylamide (UCL 1421-H)  1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid n-butylamide (UCL 1397-H)  1- (2 (S) -aminobutyryl) -2- (S) -indolinecarboxylic acid n -propylamide (UCL 1413-H)  1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid methylamide (UCL 1414-H)  1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid ethylamide (UCL 1415-H)  1- (2 (S) -aminobutyryl) -2 (R 1 S) - (5-methoxy) indole carboxylic acid n-butylamide (UCL 1486-H)  1- (2 (S) -aminobutyryl, -2 (R / S) - (6-methoxy) indoline carboxylic acid n-butylamide (UCL 1546-H)  1- (2 (S) -aminobutyryl) -2 (R / S) - (5-fluoro) indoline carboxylic acid n-butylamide (UCL 1534-H)  1- (2 (S) -aminobutyryl) -2 (RIS) - (S-benzyloxy) indoline carboxylic acid n-butylamide (UCL 1535-H)  1- (2 (S) -aminobutyryl) -2 (S) - [(3aS, 7aS) -perhydro] -indolinecarboxylic acid n-butylamide (UCL 1442-H)  2- (2 (S) -aminobutyryl) -1 (R / S) -isoindoline carboxylic acid n-butylamide (UCL 1466-H)  1- (2 (S) -aminobutyryl) -2-hexylpyrrolidineamide (UCL 1419-H)  1- (2 (S) -aminobutyryi) -2 - [(1-oxo) hexy] pyrrolidineamide (UCL 1420-H)  1- (2-Amino-2-methylpropionyl) -L-proline n-butylamide (UCL 1424-A)  1- (2 (S) -aminobutyryl) -2 (R / S) -perhydroazepine carboxylic acid n-butylamide (UCL 1482-H)  1- (2 (S) -aminobutyryl) -2 (S) -indolinylmethyl-N-butylamine (UCL 1446-A2) and their corresponding salts or hydrates.  32 / A compound according to claim 17, characterized in that it is chosen from the group comprising  1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid n-butylamide (UCL 1397-H)  1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid n -propylamide (UCL 1413-H)  1- (2 (S) -aminobutyryl) -2 (S) -indolinecarboxylic acid ethylamide (UCL 1415-H)  1- (2 (S) -aminobutyryl) -2 (R / S) - (5-methoxy) indoline carboxylic acid n-butylamide (UCL 1486-H)  1- (2 (S) -aminobutyryl) -2 (R 1 S) - (5-fluoro) indoline carboxylic acid n-butylamide (UCL 1534-H) and their corresponding salts or hydrates.  33 / Process for the preparation of a compound of general formula (I) given above in which  R1 represents a linear or branched C1-C4 alkyl group  R2 represents a hydrogen  n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents - (CH2) 2-, -CH2S-, -CH2-; and  R4 represents CO-NH-R5, COR7  where Rs is hydrogen or a linear or branched C1-C6 alkyl group, - (CH2) 3-S-CH3, -CH2Ph and R7 represents NC5H10, CH2C6H11  characterized in that it comprises  i) the formation of a compound of formula (III)

 wherein R1 and R2 have the meanings given above and X represents a protecting group, from a compound of formula (II)

 ii) amidation of the acid function of the compound (III) with the appropriate amine R5NH2 or R7H where Rs and R7 have the meanings given above, to form the derivative (IV)  esterified on its acid function by a group Y and in which X, R1 and R2 have the meanings given above, by reaction with L-proline  iii) removing the X group of the derivative (IV), to obtain the desired compound (I).  protected on its primary amine function by group X  34 / Process for the preparation of a compound of general formula (I) given above in which  R1 represents a linear or branched C1-C4 alkyl group  R2 represents a hydrogen or a methyl group  - n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents a group - (CH2) 2-; and  R4 represents a CO-NH-R5 group where Rs represents a hydrogen or a linear or branched C1-C6 alkyl group;  characterized in that it comprises  i) forming a compound of formula (IV)

 wherein R1, R2 and R5 have the meanings given above and X represents a protecting group, from a compound of formula (II)

 esterified to its acid function using a group Y and in which R1, R2 and X have the meanings given above, by reaction with a prolineamide of formula (V)  ii) removal of the X protecting group of the compound (IV), to obtain the desired compound (I).  in which Rs has the meaning given above  35 / Process for the preparation of a compound of general formula (I) given above in which  R1 represents a linear or branched C1-C4 alkyl group  R2 represents a hydrogen  n = 0 and m = 1 and R3 represents a group -CH2-CH (cis.F) -,

 i) the preparation of a compound of formula (X)  characterized in that it comprises  R4 represents a group -CH2OH, CO-NH-R5 where Rs represents a hydrogen or a linear or branched C1-C6 alkyl group;

 with a compound of formula (IX)

 in which R 1, R 2, R 3 and R 4 have the meanings given above and X represents a protecting group, by reaction of a compound of formula (VIII)  ii) removing the X group of the compound (X), to form the desired compound (I).  in which R1, R2, R4 and X have the meanings given above  36 / Process for the preparation of a compound of general formula (I) given above in which  R1 represents a linear or branched C1-C4 alkyl group  R2 represents a hydrogen  n = 1 and m = 0 and R3 represents a group -CH (CH2Ph) CH2-, - (CH2) 3- or

 i) obtaining a compound (XII)  characterized in that it comprises  R4 represents a CO-NH-R5 group where Rs represents a hydrogen or a linear or branched C1-C6 alkyl group;

 with the compound of formula (XI)

 protected on its primary amine function by a protective group X and in which R1, R2 and R3 have the meanings given above, by reaction of the compound of formula (IX)

 ii) hydrolyzing the ester function of the compound (XII) thus obtained to form the compound of formula (XIII)  in which R1, R2 and R3 have the meanings given above and X represents a protective group

 iii) amidation of the acid function of the compound (XIII) using the appropriate amine R5NH2 to form the derivative of formula (XIV)  in which R1, R2, R3 and X have the meanings given above  iv) removing the X protecting group of the compound (XIV) to form the desired compound (I).  wherein R1, R2, R3, Rs and X have the meanings given above  37 / Process for the preparation of a compound of general formula (I) given above, in which  - R1 represents a methyl group  R2 represents a hydrogen  - n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents the group - (CH2) 2-; and  R4 represents a CO-NH-R5 group in which Rs represents the unit

 i) amidation of the acid function of the compound of formula (XIX)  characterized in that it comprises

 to form the derivative (XX)

 protected on its primary amine function by a protective group X and in which R1 and R2 have the meanings given above, using the amine (XVIII)  ii) removing the X protecting group of the compound (XX) to obtain the desired compound (I).  in which R1, R2, Rs and X have the meanings given above  38. The process as claimed in claim 37, characterized in that the amine (XVIII) is prepared.  i) by reaction of isopropylboronic acid with (+) - pinanediol in ether, to obtain the derivative (XV)

 ii) by reacting the compound (XV) with butyllithium and methylene chloride and then with zinc chloride ZnCl 2 in tetrahydrofuran, at low temperature, to form the derivative (XVI)

 iii) substitution of the chlorine of the compound (XVI) with butyllithium and hexamethyldisilazane in tetrahydrofuran at low temperature, to form the compound (XVII)  iv) releasing the amine function of the compound (XVII) to obtain the desired compound (XVIII).  39 / Process for the preparation of a compound of general formula (I) given above in which  - R1 represents a methyl group  R2 represents a hydrogen  - n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents a group - (CH2) 2-; and  R4 represents a CO-NH-R5 group where R5 represents (CH3) 2 -CH-BF2  characterized in that it comprises  i) the elimination of the pinane unit of the compound of formula (I) wherein R 1, R 2, n, m R 3 and R 4 have the meanings given above and R 5 represents

 by action of boron chloride sorting in methylene chloride followed by hydrolysis, to obtain the derivative (XXI)  ii) reacting the compound (XXI) with hydrofluoric acid to form the desired compound (I).  40 / Process for the preparation of a compound of general formula (I) given above in which  R1 represents a linear or branched C1-C4 alkyl group  R2 represents a hydrogen  - n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents - (CH2) 2-; and  - R4 represents the group n.C6Hl3  characterized in that it comprises  i) the reaction of a compound (XXV)

 with the compound of formula (IX)

 wherein R1 and R2 have the meanings given above and X represents a protecting group, to form the derivative of formula (XXVI)

 ii) hydrogenating the double bond of the compound (XXVI) thus obtained to form the compound of formula (XXVII)  in which R1, R2 and X have the meanings given above  (XXVII) to obtain the desired compound (I).  iii) elimination of the protective group X of the compound  wherein R1, R2 and X have the aforementioned meanings  41. The process as claimed in claim 40, wherein the compound (XXV) is prepared.  i) by reaction of the methyl ester of L-proline  (XXII)

 protected on its amine function by a protecting group X, with diisobutylaluminum hydride in toluene at low temperature, to form L-proline (XXIII)

 ii) by reacting the protected L-proline (XXIII) on its heterocyclic amine function with a protecting group X, with triphenylpentylphosphonium bromide in the presence of butyllithium in toluene, to form the derivative (XXIV)  iii) removal of the protective group X of the compound (XXIV) thus obtained.  42 / Process for the preparation of a compound of formula (I) given above, in which  R1 represents a linear or branched C1-C4 alkyl group  R2 represents a hydrogen  - n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents a group - (CH2) 2-; and  R4 represents a group CO-R7 where R7 represents the group n.C5H11  characterized in that it comprises  i) obtaining a compound of formula (XXX)

 with the compound of formula (IX)

 wherein R1 and R2 have the meanings given above and X is a protecting group, by reaction of the compound (XXIX)  ii) removing the X protecting group of the obtained compound (XXX), to form the desired compound (I).  wherein R1, R2 and X have the meanings given above;  43 / A method according to claim 42, characterized in that the compound (XXIX) is prepared by hydrogenation of the corresponding hexanone (XXVIII)

 (XXVIII) of L-proline, obtained by action of pentyllithium in the ether, on the L-proline protected on its amine function by an X group.  44 / Process for the preparation of a compound of general formula (I) given above, in which  - R1 and R2 each represent a methyl group  - n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents a group - (CH2) 2-; and  R4 represents a CO-NH-R5 group where R5 represents a linear or branched C1-C6 alkyl group;  characterized in that it comprises  i) the reaction of the appropriate L-prolineamide of

 with the acid of formula (IX)

 in which R1, R2 and R5 have the meanings given above and X is a protecting group, to form the derivative of formula (IV)  ii) removing the protective group X of the compound (IV) to obtain the compound (I) sought.  wherein R1, R2, Rs and X have the above meanings  45 / Process for the preparation of a compound of general formula (I) given above, in which  - R1 represents a hydrogen  - R2 is different from hydrogen  - n = 0 and m = 1 and R3 represents a pattern

 - R4 represents the group CH2-NH (n.C4H9) characterized in that it comprises i) the formation of a compound of formula (XXXII)

 from the corresponding amide of formula (XXXI)

 ii) protecting the secondary amine function of the compound (XXXII) thus formed by a group X ', to obtain the compound (XXXIII)  in which X represents a protecting group

 iii) selective deprotection of the heterocyclic amine by removal of the X group of the compound (XXXIII), to form the compound (XXXIV)

 iv) the reaction of the compound (XXXIV) with the compound of formula (IX)

 wherein R1, R2 and X have the above meanings to form a compound of formula (XXXV)  v) removing the X and X 'protecting groups of the compound (XXXV) to form the desired compound (I).  in which R1, R2 X and X 'have the meanings indicated above  46 / Process for the preparation of a compound of general formula (I) given above, in which  R1 represents a linear or branched C1-C4 alkyl group  R2 represents a hydrogen  - n = 0 or 1 and m = 0 or 1 with n different from m and R3 represents a group - (CH2) 4-; and  - R4 represents a CO-NH-R5 group where R5 represents a linear or branched C1-C6 alkyl group  characterized in that it comprises  i) formation of the compound of formula (XXXVI)

 from the compound of formula (IX)

 ii) reaction of the compound (XXXVI) with 2- (RSS) -perhydroazepine acid, to form the derivative (XXXVII)  wherein R1 and R2 have the meanings given above and X is a protecting group, using 3-hydroxy-1,2,3-benzotriazine-4 (3H) -one in the presence of dicyclohexylcarbodiimide (DCC) in tetrahydrofuran

 iii) amidation of the acid function of the compound (XXXVII) thus obtained, using the appropriate amine R5NH2, to form the derivative of formula (XXXVIII)  wherein R1, R2 and X have the aforementioned meanings  iv) removing the X protecting group of the compound (XXXVIII) to obtain the desired compound (I).  wherein R1, R2 and X have the aforementioned meanings  47 / Process for the preparation of a compound of general formula (I) given above, in which  R1 represents a linear or branched C1-C4 alkyl group  R2 represents a hydrogen  - n = 0 and m = 1 and R3 represents the motive

 where R6 is OCR3, OCH2Ph, F; and  - R4 represents a CO-NH-R5 group where R5 represents a linear or branched C1-C6 alkyl group  characterized in that it comprises  i) formation of the amide of formula (XXXXIV)

 from the ester of formula (XXXXIII)

 ii) the reaction of the compound (XXXXIV) with the compound of formula (IX)  wherein R6 has the meaning given above, by reaction with the appropriate amine R5NH2

X represents a protecting group, to form the compound of formula (XXXXV)  wherein R1 and R2 have the aforementioned meanings and  iii) removing the X group to form the desired compound (I).  in which R1, R2, Rg, R6 and X have the meanings given above  48 / Process according to Claim 47, characterized in that the methyl ester (XXXXIII) is prepared  i) by the action of sodium nitrite in the presence of hydrochloric acid, on the compound of formula (XXXIX)

 to form the compound of formula (XXXX)

 ii) by adding ethyl 2-methylacetoacetate to the compound (XXXX) thus obtained, in the presence of sodium nitrite in ethanol, to form the compound of formula (XXXXI)  in which R6 has the meaning indicated above

 iii) by cyclization in acidic medium, to form the ethyl ester (XXXXII)  in which K6 has the above mentioned  iv) by exchange from the ethyl ester (XXXXII) thus obtained, in the presence of magnesium in methanol.  49 / Use of a compound of general formula (I) according to any one of claims 7 to 32 as a medicament for the treatment in humans or animals of disorders or conditions related to the inactivation of endogenous neuropeptides .  50 / Use according to claim 49, characterized in that the disorders or conditions are related to the inactivation of cholecystokinin (CCK).  51 / Use according to claim 50, characterized in that the disorders or conditions are disorders of food intake, mood, cognitive or motor, including anorexia, schizophrenia, Parkinson's disease and depressions as well as gastrointestinal transit disorders such as irritable bowel syndrome.  52 / Use of a compound of general formula (I) according to any one of claims 7 to 32 for the preparation of a medicament for the treatment in humans or animals of disorders or conditions related to inactivation of endogenous neuropeptides.  53 / Use according to revendicaiton 52, characterized in that the disorders or conditions are related to the inactivation of cholecystokinin (CCK).  54 / Use according to claim 53, characterized in that the disorders or conditions are disorders of food intake, mood, cognitive or motor, including anorexia, schizophrenia, Parkinson's disease and depressions as well as gastrointestinal transit disorders such as irritable bowel syndrome.  A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 7 to 30 in a pharmaceutically acceptable excipient.  56 / Pharmaceutical composition according to claim 55, characterized in that it comprises a compound according to claim 31.  57 / Pharmaceutical composition according to claim 55, characterized in that it comprises a compound according to claim 32.  58 / The use according to claim 49, characterized in that a compound of formula (I) in which R 1 represents a CH 2 CH 3, (CH 2) 2 CH 3, (CH 2) 3 CH 3, CH (CH 3) 2, CH 2 CH group is used. (CH 3) 2, CH (CH 3) CH 2 CH 3, R 2 is hydrogen, R 3 is the divalent radical - (CH 2) 2 and R 4 is the amide group CO-NH-R 5, where R 5 is hydrogen.  59 / Use according to claim 52, characterized in that one uses a compound of formula (I) wherein R1 represents a group CH2CH3, (CH2) 2CH3, (CH2) 3CH3, CH (CH3) 2, CH2CH (CH 3) 2, CH (CH 3) CH 2 CH 3, R 2 represents hydrogen, R 3 represents the divalent radical - (CH 2) 2 and R 4 represents the amide group CO-NH-R 5, R 8 being hydrogen.